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Sample records for a549 lung cancer

  1. Exosomes: Decreased Sensitivity of Lung Cancer A549 Cells to Cisplatin

    Xia Xiao; Shaorong Yu; Shuchun Li; Jianzhong Wu; Rong Ma; Haixia Cao; Yanliang Zhu; Jifeng Feng

    2014-01-01

    Exosomes are small extracellular membrane vesicles of endocytic origin released by many cells that could be found in most body fluids. The main functions of exosomes are cellular communication and cellular waste clean-up. This study was conducted to determine the involvement of exosomes in the regulation of sensitivity of the lung cancer cell line A549 to cisplatin (DDP). When DDP was added to A549 cells, exosomes secretion was strengthened. Addition of the secreted exosomes to other A549 cel...

  2. Klotho inhibits growth and promotes apoptosis in human lung cancer cell line A549

    Zhao Weihong

    2010-07-01

    Full Text Available Abstract Background Klotho, as a new anti-aging gene, can shed into circulation and act as a multi-functional humoral factor that influences multiple biological processes. Recently, published studies suggest that klotho can also serve as a potential tumor suppressor. The aim of this study is to investigate the effects and possible mechanisms of action of klotho in human lung cancer cell line A549. Methods In this study, plasmids encoding klotho or klotho specific shRNAs were constructed to overexpress or knockdown klotho in vitro. A549 cells were respectively treated with pCMV6-MYC-KL or klotho specific shRNAs. The MTT assay was used to evaluate the cytotoxic effects of klotho and flow cytometry was utilized to observe and detect the apoptosis of A549 cells induced by klotho. The activation of IGF-1/insulin signal pathways in A549 cells treated by pCMV6-MYC-KL or shRNAs were evaluated by western blotting. The expression levels of bcl-2 and bax transcripts were evaluated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR. Results Overexpression of klotho reduced the proliferation of lung cancer A549 cells, whereas klotho silencing in A549 cells enhanced proliferation. Klotho did not show any effects on HEK-293 cells. Klotho overexpression in A549 cells was associated with reduced IGF-1/insulin-induced phosphorylation of IGF-1R (IGF-1 receptor/IR (insulin receptor (P P P P P Conclusions Klotho can inhibit proliferation and increase apoptosis of A549 cells, this may be partly due to the inhibition of IGF-1/insulin pathways and involving regulating the expression of the apoptosis-related genes bax/bcl-2. Thus, klotho can serve as a potential tumor suppressor in A549 cells.

  3. Xylitol induces cell death in lung cancer A549 cells by autophagy.

    Park, Eunjoo; Park, Mi Hee; Na, Hee Sam; Chung, Jin

    2015-05-01

    Xylitol is a widely used anti-caries agent that has anti-inflammatory effects. We have evaluated the potential of xylitol in cancer treatment. It's effects on cell proliferation and cytotoxicity were measured by MTT assay and LDH assay. Cell morphology and autophagy were examined by immunostaining and immunoblotting. Xylitol inhibited cell proliferation in a dose-dependent manner in these cancer cells: A549, Caki, NCI-H23, HCT-15, HL-60, K562, and SK MEL-2. The IC50 of xylitol in human gingival fibroblast cells was higher than in cancer cells, indicating that it is more specific for cancer cells. Moreover, xylitol induced autophagy in A549 cells that was inhibited by 3-methyladenine, an autophagy inhibitor. These results indicate that xylitol has potential in therapy against lung cancer by inhibiting cell proliferation and inducing autophagy of A549 cells. PMID:25650339

  4. Wnt/β-catenin signaling regulates cancer stem cells in lung cancer A549 cells

    Wnt/β-catenin signaling plays an important role not only in cancer, but also in cancer stem cells. In this study, we found that β-catenin and OCT-4 was highly expressed in cisplatin (DDP) selected A549 cells. Stimulating A549 cells with lithium chloride (LiCl) resulted in accumulation of β-catenin and up-regulation of a typical Wnt target gene cyclin D1. This stimulation also significantly enhanced proliferation, clone formation, migration and drug resistance abilities in A549 cells. Moreover, the up-regulation of OCT-4, a stem cell marker, was observed through real-time PCR and Western blotting. In a reverse approach, we inhibited Wnt signaling by knocking down the expression of β-catenin using RNA interference technology. This inhibition resulted in down-regulation of the Wnt target gene cyclin D1 as well as the proliferation, clone formation, migration and drug resistance abilities. Meanwhile, the expression of OCT-4 was reduced after the inhibition of Wnt/β-catenin signaling. Taken together, our study provides strong evidence that canonical Wnt signaling plays an important role in lung cancer stem cell properties, and it also regulates OCT-4, a lung cancer stem cell marker.

  5. [Grape seed proanthocyanidins inhibits the invasion and migration of A549 lung cancer cells].

    Zhou, Yehan; Ye, Xiufeng; Shi, Yao; Wang, Ke; Wan, Dan

    2016-02-01

    Objective To explore the effect of grape seed proanthocyanidins (GSPs) on the invasion and migration of A549 lung cancer cells and the underlying mechanism. Methods Trypan blue dye exclusion assay was used to determine the cytotoxic effect of varying doses of GSPs on the BEAS-2B normal human pulmonary epithelial cells. After treated with 0, 10, 20, 40, 80 μg/mL GSP, the proliferation of A549 cells was detected by MTT assay; the invasion and migration of A549 cells were determined by Transwell(TM) assay and scratch wound assay, respectively. The levels of epithelial growth factor receptor (EGFR), E-cadherin, N-cadherin in A549 cells treated with GSPs were detected by Western blotting. Results (0-40) μg/mL GSPs had no significant toxic effect on BEAS-2B cells, while 80 μg/mL GSPs had significant cytotoxicity to BEAS-2B cells. The proliferation of A549 cells was significantly inhibited within limited dosage in a dose-dependent manner, and the abilities of invasion and migration of A549 cells were also inhibited. Western blotting showed that the expression of EGFR and N-cadherin decreased, while E-cadherin increased after GSPs treatment. Conclusion GSPs could inhibit the abilities of proliferation, invasion and migration of A549 cells, which might be related to the dow-regulation of EGFR and N-cadherin and the up-regulation of E-cadherin. PMID:26927375

  6. MicroRNA-126 inhibits the proliferation of lung cancer cell line A549

    Xun Yang; Bei-Bei Chen; Ming-Hua Zhang; Xin-Rong Wang

    2015-01-01

    Objective:To study the role of microRNA-126 in the development of lung cancer.Methods:The biological function of microRNA-126 was detected using EdU assay and CCK-8 assay;the target gene of microRNA-126 was analyzed using real time RT-PCR and Western blot assay.Results: In A549 cell line, overexpression of microRNA-126 inhibits the proliferation rate; VEGF is the target gene of microRNA-126; microRNA-126 exerts its function via regulating VEGF protein level.Conclusions: microRNA-126 inhibits the proliferation in A549 cell line.

  7. Effect of Inhibiting NGAL Gene Expression on A549 Lung Cancer Cell Migration and Invasion

    Jian TANG

    2015-04-01

    Full Text Available Background and objective To detect the expression of neutrophil gelatinase-assoeiated lipocalin (NGAL in the different differentiations of lung cancer tissues and to study the mechanism of invasion of A549 cells affected by NGAL. Methods The expression of NGAL was detected by immunochemistry in lung cancer tissue and the tissue around edge of the cancer. The effect of NGAL expression on A549 cells was observed by using qRT-PCR and Western blot. The abilities of invasion and metastasis were evaluated by transwell invasion and migration assay, and cell scratch assay in vitro. The protein expression of E-cadherin, Vimentin was measured by immunofluoresence and Western blot. Results The positive expression rate of NGAL was 76.32% (58/76 in the lung cancer, 13.3% (4/30 in adjacent tissue by immunochemistry. NGAL expression levels in the lung cancer tissues were significantly higher than that in adjacent tissues. The rate of migration and invasion in NGAL-siRNA group was 60.4%±6.4% compared to 50.5%±4.4% in the control group, there was a significant difference (P<0.05. Vimentin was suppressed, and E-cadherin was upregulated when NGAL was inhibited. MMP-2 and MMP-9 decreased when NGAL was knocked down. Conclusion The expression level of NGAL is highly expressed in lung cancer. NGAL may be one of important indicators involved in lung cancer infiltrated and transferred. NGAL might be one of potential targets for lung cancer treatment.

  8. Effect of evodiamine on the proliferation and apoptosis of A549 human lung cancer cells.

    Lin, Li; Ren, Li; Wen, Liujing; Wang, Yu; Qi, Jin

    2016-09-01

    Evodia rutaecarpa is a plant, which has antitumor activity. Evodiamine is an alkaloid with antitumor activity present in E. rutaecarpa and has potential to be developed into a therapeutic antitumor agent. The present study investigated the effect of evodiamine on the proliferation of A549 human lung cancer cells and the mechanism underlying these effects. The results indicated that evodiamine significantly inhibited proliferation, induced apoptosis and the expression of reactive oxygen species, arrested the cell cycle, regulated the expression of Survivin, Bcl-2 and Cyclin B1, regulated the activity of caspase-3/8 and glutathione in tumor cells, and decreased the activity of AKT/nuclear factor‑κB (NF‑κB) and Sonic hedgehog/GLI family zinc finger 1 (SHH/GLI1) signaling pathways in A549 cells. In conclusion, the evodiamine-induced inhibition of the proliferation of A549 lung cancer cells may be attributable to its ability to promote oxidative injury in the cells, induce apoptosis, arrest the cell cycle and regulate the AKT/NF‑κB and SHH/GLI1 signaling pathways, subsequently controlling the expression of tumor‑associated genes. PMID:27485202

  9. Epithelial mesenchymal transition of non-small-cell lung cancer cells A549 induced by SPHK1

    Min Ni; Xiao-Lei Shi; Zhi-Gang Qu; Hong Jiang; Zi-Qian Chen; Jun Hu

    2015-01-01

    Objective:To explore the effect and molecular mechanism ofSPHK1 in the invasion and metastasis process of non-small-cell lung cancer cells(A549).Methods:Recombinant retrovirus was used to mediate the production ofA549/vector,A549/SPHK1,A549/scramble, andA549/SPHK1/RNAi that stably expressed or silencedSPHK1.The invasion and migration capacities of A549 cells overexpressing or silencingSPHK1 were determined usingTranswell invasion assay and scratch wound repair experiment.The protein and mRNA expression levels ofE-cadherin, fibronectin, vimentin inA549/vector,A549/SPHK1,A549/scramble,A549/SPHK1/RNAi were detected withWestern blot(WB) and quantitativePCR(QPCR) methods, respectively.Results:Transwell invasion assay and scratch wound repair experiments showed that over-expression of SPHK1 obviously enhanced the invasion and migration capacities ofA549 cells.WB andQPCR detection results showed that, the expression ofE-cadherin(a molecular marker of epithelial cells) and fibronectin, vimentin(molecular markers of mesenchymal cells) inA549 cells was upregulated after overexpression ofSPHK1; whileSPHK1 silencing significantly reduced the invasion and metastasis capacities ofA549cells, upregulated the expression of molecular marker of epithelial cells, and downregulated the expression of molecular marker of mesenchymal cells. Conclusions:SPHK1 promotes epithelial mesenchymal transition of non-small-cell lung cancer cells and affects the invasion and metastasis capacities of these cells.

  10. ANTICANCER ACTIVITY OF OSCILLATORIA TEREBRIFORMIS CYANOBACTERIA IN HUMAN LUNG CANCER CELL LINE A549

    S.Mukund

    2014-04-01

    Full Text Available Purpose: To evaluate the anti-cancer properties of the cyanobacterial extract of Oscillatoria terebriformis Methods: The extract was tested in Human lung cancer cell lines and examined for its effect on cell viability, nuclear morphology and sub-G1 formation. Cell viability was determined by micro culture tetrazolium technique (MTT, nuclear morphology investigated using 4’-6-diamidino-2-phenylindole (DAPI staining technique, and apoptosis assay using DNA fragmentation. Results: The results showed decreasing cell viability in a concentration-dependent manner. Altered cell morphology after treatment with the extract demonstrated that cells experienced apoptosis. Conclusion: The data demonstrate that Oscillatoria Terebriformis extract induced apoptosis in Human lung cancer A549 cells, and therefore, has a potential as an anti-cancer agent.

  11. COPD promotes migration of A549 lung cancer cells: the role of chemokine CCL21

    Kuźnar-Kamińska B

    2016-05-01

    Full Text Available Barbara Kuźnar-Kamińska,1 Justyna Mikuła-Pietrasik,2 Patrycja Sosińska,2 Krzysztof Książek,2 Halina Batura-Gabryel1 1Department of Pulmonology, Allergology and Respiratory Oncology, 2Department of Pathophysiology, Poznań University of Medical Sciences, Poznań, Poland Abstract: Patients with COPD develop lung cancer more frequently than healthy smokers. At the same time, molecular mediators promoting various aspects of cancer cell progression are still elusive. In this report, we examined whether COPD can be coupled with increased migration of non-small-cell lung cancer cells A549 and, if so, whether this effect may be related to altered production and activity of chemokines CCL21, CXCL5, and CXCL12. The study showed that the migration of A549 cells through the polycarbonate membrane and basement membrane extract toward a chemotactic gradient elicited by serum from patients with COPD was markedly higher as compared with serum from healthy donors. The concentration of CCL21 and CXCL12, but not CXCL5, in serum from patients with COPD was also increased. Experiments in which CCL21- and CXCL12-dependent signaling was blocked revealed that increased migration of the cancer cells upon treatment with serum from patients with COPD was mediated exclusively by CCL21. Collectively, our results indicate that COPD may contribute to the progression of lung cancer via CCL21-dependent intensification of cancer cell migration. Keywords: chemokines, COPD, lung cancer, migration

  12. Enhancement of radiation sensitivity by erlotinib and celecoxib in A549 human lung cancer cell line

    Objective: To investigate the role of epidermal growth factor receptor and cyclooxygenase-2 pathways in the erlotinib and celecoxib enhanced radiation sensitivity in A549 human lung cancer cell line. Methods: IC20 of erlotinib and celecoxib on in A549 human lung cancer cells was measured by MTT assay, Clonogenic assays were used to evaluate the antitumor effects of the drugs and X-irradiation. Flow cytometry was used to assess the apoptosis and cell cycle alteration, and Western blot was used for the detection of Akt and phospho-Akt.Results Both erlotinib and celecoxib could inhibit the proliferation of A549 cells in vitro in a dose-dependent manner and their values of IC20 were (5.15 ± 0.14) and (40.32 ± 1.26) μmol/L, respectively. For radiation survival,the values of Dq, D0, SF2 of the combination of two drugs were lower than those of either drug (t=6.62, P<0.05). The SER of celecoxib, erlotinib and their combination were 1.299, 1.503 and 2.217, respectively. Flow cytometry assay showed that both celecoxib and erlotinib could enhance radiation-induced G0/G1 arrest, reduce the cell number in S phase, and enhance radiation-induced apoptosis, especially for the combination of drugs. Western blot assay showed that the expressions of Akt protein were similar in all groups. However, pAkt expression was suppressed by erlotinib and celecoxib, but promoted by radiation. pAkt had the lowest expression in the radiated cells with the treatment of two drugs (t=4.89, P<0.05). Conclusions: The erlotinib and/or celecoxib could enhance radiosensitivity probably by increasing cell apoptosis and reducing the number of S-phase cells with low radiosensitivity. (authors)

  13. Apoptosis inducing ability of silver decorated highly reduced graphene oxide nanocomposites in A549 lung cancer

    Khan M

    2016-03-01

    Full Text Available Merajuddin Khan,1 Mujeeb Khan,1 Abdulhadi H Al-Marri,1 Abdulrahman Al-Warthan,1 Hamad Z Alkhathlan,1 Mohammed Rafiq H Siddiqui,1 Vadithe Lakshma Nayak,2 Ahmed Kamal,2 Syed F Adil1 1Department of Chemistry, College of Science, King Saud University, Riyadh, Kingdom of Saudi Arabia; 2Department of Medicinal Chemistry and Pharmacology, CSIR – Indian Institute of Chemical Technology, Hyderabad, India Abstract: Recently, graphene and graphene-based materials have been increasingly used for various biological applications due to their extraordinary physicochemical properties. Here, we demonstrate the anticancer properties and apoptosis-inducing ability of silver doped highly reduced graphene oxide nanocomposites synthesized by employing green approach. These nano­composites (PGE-HRG-Ag were synthesized by using Pulicaria glutinosa extract (PGE as a reducing agent and were evaluated for their anticancer properties against various human cancer cell lines with tamoxifen as the reference drug. A correlation between the amount of Ag nanoparticles on the surface of highly reduced graphene oxide (HRG and the anticancer activity of nanocomposite was observed, wherein an increase in the concentration of Ag nanoparticles on the surface of HRG led to the enhanced anticancer activity of the nanocomposite. The nanocomposite PGE-HRG-Ag-2 exhibited more potent cytotoxicity than standard drug in A549 cells, a human lung cancer cell line. A detailed investigation was undertaken and Fluorescence activated cell sorting (FACS analysis demonstrated that the nanocomposite PGE-HRG-Ag-2 showed G0/G1 phase cell cycle arrest and induced apoptosis in A549 cells. Studies such as, measurement of mitochondrial membrane potential, generation of reactive oxygen species (ROS and Annexin V-FITC staining assay suggested that this compound induced apoptosis in human lung cancer cells. Keywords: plant extract, graphene/silver nanocomposites, anticancer, apoptosis

  14. Screening Metastasis-associated Genes from Anoikis Resistant A549 Lung Cancer Cells by Human Genome Array

    Xiaoping WANG

    2010-01-01

    Full Text Available Background and objective As a barrier to metastases, cells normally undergo apoptosis after they lose contact with their extra cellular matrix (ECM. This process has been termed “anoikis”. Tumour cells that acquire malignant potential have developed mechanisms to resist anoikis and thereby survive after detachment from their primary site while traveling through the lymphatic and circulatory systems. This “anoikis resistance” is considered the first step to tumor metastases. The aim of this study was to screen metastasis-associated genes from anoikis resistant and adherent growth A549 lung cancer cell by Human Genome Array. Methods Establish anoikis resistant A549 lung cancer cell lines by using poly-hydroxyethyl methacrylate resin processed petri dishes, which causes cell free from adherent. The different expressed gene between anoikis resistant A549 cell and adherent growth A549 cell was tested using human V2.0 whole-genome oligonucleotide microarray, a product of Capitalbio Corporation, Beijing. Screen metastasis-associated genes. Results 745 different expressed genes were screened, including 63 highly metastasis-associated genes. Conclusion The successfully established anoikis resistant A549 cell lines and screened different expressed genes provide us basis for further research on metastasis of lung cancer.

  15. Growth arrest and apoptosis via caspase activation of dioscoreanone in human non-small-cell lung cancer A549 cells

    Hansakul, Pintusorn; Aree, Kalaya; Tanuchit, Sermkiat; Itharat, Arunporn

    2014-01-01

    Background Dioscoreanone (DN) isolated from Dioscorea membranacea Pierre has been reported to exert potent cytotoxic effects against particular types of cancer. The present study was carried out to investigate the cytotoxicity of DN against a panel of different human lung cancer cell lines. The study further examined the underlying mechanisms of its anticancer activity in the human lung adenocarcinoma cell line A549. Methods Antiproliferative effects of DN were determined by SRB and CFSE assa...

  16. The Effects of Davallic Acid from Davallia divaricata Blume on Apoptosis Induction in A549 Lung Cancer Cells

    Tsu-Liang Chang

    2012-11-01

    Full Text Available Traditional or folk medicinal herbs continue to be prescribed in the treatment of various diseases and conditions in many cultures. Recent scientific efforts have focused on the potential roles of extracts of traditional herbs as alternative and complementary medications for cancer treatment. In Taiwan, Davallia divaricata Blume has been traditionally employed in folk medicine for therapy of lung cancer, davallic acid being the major active compound of D. divaricata Blume. In this study, we investigated the inhibitory activity of davallic acid on the proliferation of A549 lung cancer cells. Davallic acid was extracted from D. divaricata Blume, and its effects on cell viability, cell cycle distribution, ROS level, and apoptotic protein expression in A549 cells were determined. Davallic acid significantly induced reactive oxygen species (ROS generation as well as caspase-3, -8, and -9 activation, thereby repressing A549 cell growth and elevating apoptotic activity. Since lung cancer has a high incidence of recurrence, these results indicate that davallic acid may have the potential to be a natural anti-lung cancer compound, and may provide a basis for further study of its use in combating cancer.

  17. Inhibitory Effects of Natural Compound Alantolactone on Human Non-small Cell Lung Cancer A549 Cells

    ZONG Min-ru; ZHAO Ying-hao; ZHANG Kun; YANG Long-fei; ZHENG Yong-chen; HE Cheng-yan

    2011-01-01

    Alantolactone is a natural compound identified from the roots of Inula helenium L. that has multiple bio-activities. We examined its inhibitory effects on human non-small cell lung cancer(NSCLC) A549 cells. The antiproliferative effect of alantolactone on A549 cells was investigated via MTT[3'-(4,5dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide]assay and its apoptosis-inducing effect was determined by Hoechst staining and flow cytometry. We found that alantolactone significantly inhibited the proliferation of A549 cells and induced morphological changes typical for apoptosis. Flow cytometry analysis indicates dose-dependent cell cycle retardation at G0/G1 and S stages. The results indicate that alantolactone could be an attractive small-molecular natural compound for further development as a therapeutic drug against NSCLC.

  18. CDK-associated Cullin 1 promotes cell proliferation with activation of ERK1/2 in human lung cancer A549 cells

    Highlights: •CDK-associated Cullin 1 (CAC1) expression increases in human lung carcinoma. •CAC1 promotes the proliferation of lung cancer A549 cells. •CAC1 promotes human lung cancer A549 cell proliferation with activation of ERK1/2. -- Abstract: Lung cancer is one of the most common causes of cancer-related death in the world, but the mechanisms remain unknown. In this study, we investigated the expression of CDK-associated Cullin 1 (CAC1) in lung cancer, the effect of CAC1 on the proliferation of human lung cancer A549 cells, and the activation of signaling pathways of mitogen-activated protein kinases (MAPKs). Results showed that CAC1 expression was higher levels in human lung carcinoma than normal lung tissue, and CAC1 siRNA reduced the proliferation of lung cancer A549 cells by decreasing cell activity and cell division in vitro. The proportion of cells treated with CAC1 siRNA increased in the G1 phase and decreased in the S and G2/M phase, indicative of G1 cell cycle arrest. Furthermore, the proportions of early/late apoptosis in lung cancer A549 cells were enhanced with CAC1 siRNA treatment. It was also found that activation of extracellular signal-regulated protein kinase (ERK) and p38 signaling pathways were involved in the proliferation of A549 cells. After CAC1 siRNA treatment, p-ERK1/2 levels decreased, and meanwhile p-p38 level increased, A549 cell proliferation increased when ERK1/2 signaling is activated by PMA. Our findings demonstrated that CAC1 promoted the proliferation of human lung cancer A549 cells with activation of ERK1/2 signaling pathways, suggesting a potential cure target for treatment of human lung cancer

  19. CDK-associated Cullin 1 promotes cell proliferation with activation of ERK1/2 in human lung cancer A549 cells

    Chen, Tian Jun [Respiratory Department, The First Affiliated Hospital, Xi’an Jiaotong University College of Medicine, Xi’an 710061 (China); Gao, Fei [Hua-shan Central Hospital of Xi’an, Xi’an 710043 (China); Yang, Tian; Thakur, Asmitanand; Ren, Hui; Li, Yang; Zhang, Shuo; Wang, Ting [Respiratory Department, The First Affiliated Hospital, Xi’an Jiaotong University College of Medicine, Xi’an 710061 (China); Chen, Ming Wei, E-mail: xjtucmw@163.com [Respiratory Department, The First Affiliated Hospital, Xi’an Jiaotong University College of Medicine, Xi’an 710061 (China)

    2013-07-19

    Highlights: •CDK-associated Cullin 1 (CAC1) expression increases in human lung carcinoma. •CAC1 promotes the proliferation of lung cancer A549 cells. •CAC1 promotes human lung cancer A549 cell proliferation with activation of ERK1/2. -- Abstract: Lung cancer is one of the most common causes of cancer-related death in the world, but the mechanisms remain unknown. In this study, we investigated the expression of CDK-associated Cullin 1 (CAC1) in lung cancer, the effect of CAC1 on the proliferation of human lung cancer A549 cells, and the activation of signaling pathways of mitogen-activated protein kinases (MAPKs). Results showed that CAC1 expression was higher levels in human lung carcinoma than normal lung tissue, and CAC1 siRNA reduced the proliferation of lung cancer A549 cells by decreasing cell activity and cell division in vitro. The proportion of cells treated with CAC1 siRNA increased in the G1 phase and decreased in the S and G2/M phase, indicative of G1 cell cycle arrest. Furthermore, the proportions of early/late apoptosis in lung cancer A549 cells were enhanced with CAC1 siRNA treatment. It was also found that activation of extracellular signal-regulated protein kinase (ERK) and p38 signaling pathways were involved in the proliferation of A549 cells. After CAC1 siRNA treatment, p-ERK1/2 levels decreased, and meanwhile p-p38 level increased, A549 cell proliferation increased when ERK1/2 signaling is activated by PMA. Our findings demonstrated that CAC1 promoted the proliferation of human lung cancer A549 cells with activation of ERK1/2 signaling pathways, suggesting a potential cure target for treatment of human lung cancer.

  20. MicroRNA-490-3p inhibits proliferation of A549 lung cancer cells by targeting CCND1

    Highlights: • We examined the level of miR-490-3p in A549 lung cancer cells compared with normal bronchial epithelial cell line. • We are the first to show the function of miR-490-3p in A549 lung cancer cells. • We demonstrate CCND1 may be one of the targets of miR-490-3p. - Abstract: MicroRNAs (miRNAs) are small non-coding RNAs that negatively regulate the translation of messenger RNAs by binding their 3′-untranslated region (3′UTR). In this study, we found that miR-490-3p is significantly down-regulated in A549 lung cancer cells compared with the normal bronchial epithelial cell line. To better characterize the role of miR-490-3p in A549 cells, we performed a gain-of-function analysis by transfecting the A549 cells with chemically synthesized miR-490-3P mimics. Overexpression of miR-490-3P evidently inhibits cell proliferation via G1-phase arrest. We also found that forced expression of miR-490-3P decreased both mRNA and protein levels of CCND1, which plays a key role in G1/S phase transition. In addition, the dual-luciferase reporter assays indicated that miR-490-3P directly targets CCND1 through binding its 3′UTR. These findings indicated miR-490-3P could be a potential suppressor of cellular proliferation

  1. Shikonin Induces Apoptosis, Necrosis, and Premature Senescence of Human A549 Lung Cancer Cells through Upregulation of p53 Expression

    Yueh-Chiao Yeh

    2015-01-01

    Full Text Available Shikonin, a natural naphthoquinone pigment isolated from Lithospermum erythrorhizon, has been reported to suppress growth of various cancer cells. This study was aimed to investigate whether this chemical could also inhibit cell growth of lung cancer cells and, if so, works via what molecular mechanism. To fulfill this, A549 lung cancer cells were treated with shikonin and then subjected to microscopic, biochemical, flow cytometric, and molecular analyses. Compared with the controls, shikonin significantly induced cell apoptosis and reduced proliferation in a dose-dependent manner. Specially, lower concentrations of shikonin (1–2.5 μg/mL cause viability reduction; apoptosis and cellular senescence induction is associated with upregulated expressions of cell cycle- and apoptotic signaling-regulatory proteins, while higher concentrations (5–10 μg/mL precipitate both apoptosis and necrosis. Treatment of cells with pifithrin-α, a specific inhibitor of p53, suppressed shikonin-induced apoptosis and premature senescence, suggesting the role of p53 in mediating the actions of shikonin on regulation of lung cancer cell proliferation. These results indicate the potential and dose-related cytotoxic actions of shikonin on A549 lung cancer cells via p53-mediated cell fate pathways and raise shikonin a promising adjuvant chemotherapeutic agent for treatment of lung cancer in clinical practice.

  2. β-elemene reverses the drug resistance of lung cancer A549/DDP cells via the mitochondrial apoptosis pathway.

    Yao, Cheng-Cai; Tu, Yuan-Rong; Jiang, Jie; Ye, Sheng-Fang; Du, Hao-Xin; Zhang, Yi

    2014-05-01

    β-elemene (β-ELE) is a new anticancer drug extracted from Curcuma zedoaria Roscoe and has been widely used to treat malignant tumors. Recent studies have demonstrated that β-ELE reverses the drug resistance of tumor cells. To explore the possible mechanisms of action of β-ELE, we investigated its effects on cisplatin-resistant human lung adenocarcinoma A549/DDP cells. The effects of β-ELE on the growth of A549/DDP cells in vitro were determined by MTT assay. Apoptosis was assessed by fluorescence microscopy with Hoechst 33258 staining and flow cytometry with Annexin V-FITC/PI double staining. Mitochondrial membrane potential was assessed using JC-1 fluorescence probe and laser confocal scanning microscopy, and intracellular reactive oxygen species levels were measured by 2',7'-dichlorofluorescein-diacetate staining and flow cytometry. Cytosolic glutathione content was determined using GSH kits. The expression of cytochrome c, caspase-3, procaspase-3 and the Bcl-2 family proteins was assessed by western blotting. The results demonstrated that β-ELE inhibited the proliferation of A549/DDP cells in a time- and dose-dependent manner. Furthermore, β-ELE enhanced the sensitivity of A549/DDP cells to cisplatin and reversed the drug resistance of A549/DDP cells. Consistent with a role in activating apoptosis, β-ELE decreased mitochondrial membrane potential, increased intracellular reactive oxygen species concentration and decreased the cytoplasmic glutathione levels in a time- and dose-dependent manner. The combination of β-ELE and cisplatin enhanced the protein expression of cytochrome c, caspase-3 and Bad, and reduced protein levels of Bcl-2 and procaspase-3 in the A549/DDP lung cancer cells. These results define a pathway of procaspase‑3-β-ELE function that involves decreased mitochondrial membrane potential, leading to apoptosis triggered by the release of cytochrome c into the cytoplasm and the modulation of apoptosis-related genes. The reversal of drug

  3. Chemosensitization and radiosensitization of a lung cancer cell line A549 induced by a composite polymer micelle.

    Xu, Jing; Zhang, Bi-Cheng; Li, Xiang-Long; Xu, Wen-Hong; Zhou, Juan; Shen, Li; Wei, Qi-Chun

    2016-08-01

    Multidrug resistance (MDR) to Doxorubicin (DOX) remains a major obstacle to successful cancer treatment. The present study sought to overcome the MDR of lung cancer cells and achieve radiosensitization by developing a composite DOX-loaded micelle (M-DOX). M-DOX containing PEG-PCL/Pluronic P105 was prepared by the solvent evaporation method. Lung cancer cell line A549 was adopted in this study. In vitro cytotoxicity, cellular uptake behavior, subcellular distribution, and radiosensitivity were evaluated by the treatment with M-DOX, and free DOX was used as a control. A549 cells treated with M-DOX as opposed to free DOX showed greater cellular uptake as well as greater cytotoxicity. Furthermore, M-DOX reached the mitochondria and lysosome effectively after cellular uptake, and fluorescence used to track M-DOX was found to be surrounding the nucleus. Finally, colony-forming assays demonstrated that M-DOX treatment improved radiosensitization when compared to free DOX. Based on the increased cytotoxicity and radiosensitization, M-DOX could be considered as a promising drug delivery system to overcome MDR in lung cancer therapy. PMID:27585226

  4. Effects of Coptis extract combined with chemotherapeutic agents on ROS production, multidrug resistance, and cell growth in A549 human lung cancer cells

    He Chengwei

    2012-04-01

    Full Text Available Abstract Background Non–small cell lung cancer is associated with high expression of multidrug resistance (MDR proteins and low production of reactive oxygen species (ROS. Coptis extract (COP, a Chinese medicinal herb, and its major constituent, berberine (BER, have anticancer properties. This study aims to investigate the effects of COP and BER combined with chemotherapeutic agents, including fluorouracil (5-FU, camptothecin (CPT, and paclitaxel (TAX, on cell proliferation, ROS production, and MDR in A549 human non-small cell lung cancer cells. Methods A549 cells were treated with different doses of COP and BER, combined with 5-FU, CPT, and TAX. Cell viability was measured by an XTT (2,3-bis-(2-methoxy-4- nitro-5-sulfophenyl-2 H-tetrazolium-5-carboxanilide assay. Intracellular ROS levels were determined by measuring the oxidative conversion of cell permeable 2′,7′-dichlorofluorescein diacetate to fluorescent dichlorofluorescein. MDR of A549 cells was assessed by rhodamine 123 retention assay. Results Both COP and BER significantly inhibited A549 cell growth in a dose-dependent manner. Combinations of COP or BER with chemotherapeutic agents (5-FU, CPT, and TAX exhibited a stronger inhibitory effect on A549 cell growth. In addition, COP and BER increased ROS production and reduced MDR in A549 cells. Conclusion As potential adjuvants to chemotherapy for non–small cell lung cancer, COP and BER increase ROS production, reduce MDR, and enhance the inhibitory effects of chemotherapeutic agents on A549 cell growth.

  5. Inhibition of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus

    Chang HB

    2015-08-01

    Full Text Available Hong-Bin Chang,1 Bing-Huei Chen1,21Department of Food Science, 2Graduate Institute of Medicine, Fu Jen Catholic University, Taipei, TaiwanAbstract: The objectives of this study were to explore the inhibition mechanism of lung cancer cells A549 and H460 by curcuminoid extracts and nanoemulsions prepared from Curcuma longa Linnaeus. In addition, human bronchus epithelial cell line BEAS-2B (normal cell was selected for comparison. A high-performance liquid chromatography (HPLC method was developed to separate and quantify the various curcuminoids in C. longa extract, including curcumin (1,714.5 µg/mL, demethoxycurcumin (1,147.4 µg/mL, and bisdemethoxycurcumin (190.2 µg/mL. A high-stability nanoemulsion composed of Tween 80, water, and curcuminoid extract was prepared, with mean particle size being 12.6 nm. The cell cycle was retarded at G2/M for both the curcuminoid extract and nanoemulsion treatments; however, the inhibition pathway may be different. H460 cells were more susceptible to apoptosis than A549 cells for both curcuminoid extract and nanoemulsion treatments. Growth of BEAS-2B remained unaffected for both the curcuminoid extract and nanoemulsion treatments, with a concentration range from 1 to 4 µg/mL. Also, the activities of caspase-3, caspase-8, and caspase-9 followed a dose-dependent increase for both A549 and H460 cells for both the treatments, accompanied by a dose-dependent increase in cytochrome C expression and a dose-dependent decrease in CDK1 expression. Interestingly, a dose-dependent increase in cyclin B expression was shown for A549 cells for both the treatments, while a reversed trend was found for H460 cells. Both mitochondria and death receptor pathways may be responsible for apoptosis of both A549 and H460 cells.Keywords: curcuminoid extract, curcuminoid nanoemulsion, Curcuma longa Linnaeus, lung cancer cell, cell cycle, apoptosis mechanism

  6. Construction of A Eukaryotic Expression Vector Carrying the iNOS Gene and Its Effect on A549 Lung Cancer Cells

    Sujuan YE

    2012-05-01

    Full Text Available Background and objective The iNOS gene is associated with NO-mediated antitumor effects. The aims of this study are to construct a eukaryotic expression plasmid that carries the iNOS gene and to detect the expression levels and antitumor effects of the iNOS gene on A549 lung cancer cells. Methods A DNA fragment of the human iNOS coding sequence was amplified using reverse transcription polymerase chain reaction (RT-PCR. The DNA fragment was subsequently cloned into the multiple cloning sites of the eukaryotic expression vector pVAX. The recombinant plasmid was confirmed using restriction enzyme treatment, PCR, and sequencing and was then transfected into A549 lung cancer cells. The expression of the iNOS gene in the A549 lung cancer cells after transfection was verified by RT-PCR and Western blot analysis. The effects of iNOS on cell apoptosis, proliferation, and migration were identified by staining with Hoechst 3235, an MTT assay, and a scratch assay, respectively. Results The results of the restriction enzyme digestion, PCR, and sequencing verified the successful construction of the eukaryotic expression plasmid pVAX-iNOS. The iNOS gene expression level was increased in the transfected A549 cells. Further experiments also showed increased cell apoptosis among the A549 lung cancer cells transfected with pVAX-iNOS. Meanwhile, the proliferation and migration of A549 cells were significantly inhibited by the enhanced iNOS gene expression. Conclusion The recombinant eukaryotic expression vector pVAX-iNOS was successfully constructed and transfected into A549 cells. The enhanced iNOS gene expression significantly promoted cell apoptosis, whereas the proliferation and migration of A549 cells were inhibited. These findings contribute to the development of novel and effective gene therapies for lung cancer.

  7. Nanoparticles of Selaginella doederleinii leaf extract inhibit human lung cancer cells A549

    Syaefudin; Juniarti, A.; Rosiyana, L.; Setyani, A.; Khodijah, S.

    2016-01-01

    The aim of the present study is to evaluate cytotoxicity effect of nanoparticles of Selaginella doederleinii (S. doederleinii) leaves extract. S. doederleinii was extracted by maceration method using 70%(v/v) ethanol as solvent. Phytochemical content was analyzed qualitatively by using Harborne and Thin Layer Chromatography (TLC) methods. Nanoparticle extract was prepared by ionic gelation using chitosan as encapsulant agent. Anticancer activity was performed by using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The results showed that S. doederleinii contains of flavonoids. Nanoparticle of S. doederleinii leaves extract greatly inhibited A549 cells growth (cancer cells), with IC50 of 3% or 1020 μg/ml. These nanoparticles extract also inhibited the growth of Chang cells (normal cells), with IC50 of 4% or 1442 μg/ml. The effective concentration of nanoparticles extract which inhibits cancer cells without harming the normal cells is 0.5% or 167 μg/ml. Further studies are needed to obtain the concentration of nanoparticles extract which can selectively suppress cancer cells.

  8. COPD promotes migration of A549 lung cancer cells: the role of chemokine CCL21

    Kuznar-Kaminska, Barbara

    2016-01-01

    Barbara Kuźnar-Kamińska,1 Justyna Mikuła-Pietrasik,2 Patrycja Sosińska,2 Krzysztof Książek,2 Halina Batura-Gabryel1 1Department of Pulmonology, Allergology and Respiratory Oncology, 2Department of Pathophysiology, Poznań University of Medical Sciences, Poznań, Poland Abstract: Patients with COPD develop lung cancer more frequently than healthy smokers. At the same time, molecular mediators promoting various aspects of cancer cell progression are still elusive. In t...

  9. COPD promotes migration of A549 lung cancer cells: the role of chemokine CCL21

    Kuźnar-Kamińska B; Mikuła-Pietrasik J; Sosińska P; Książek K; Batura-Gabryel H

    2016-01-01

    Barbara Kuźnar-Kamińska,1 Justyna Mikuła-Pietrasik,2 Patrycja Sosińska,2 Krzysztof Książek,2 Halina Batura-Gabryel1 1Department of Pulmonology, Allergology and Respiratory Oncology, 2Department of Pathophysiology, Poznań University of Medical Sciences, Poznań, Poland Abstract: Patients with COPD develop lung cancer more frequently than healthy smokers. At the same time, molecular mediators promoting various aspects of cancer cell progression are still elusive. In this report, we e...

  10. Down-regulation of protein kinase Ceta by antisense oligonucleotides sensitises A549 lung cancer cells to vincristine and paclitaxel.

    Sonnemann, Jürgen; Gekeler, Volker; Ahlbrecht, Katrin; Brischwein, Klaus; Liu, Chao; Bader, Peter; Müller, Cornelia; Niethammer, Dietrich; Beck, James F

    2004-06-25

    Previous studies point to protein kinase C (PKC) isozyme eta as a resistance factor in cancer cells. Therefore, we investigated whether down-regulation of PKCeta with second generation antisense oligonucleotides (ODNs) would sensitise A549 human lung carcinoma cells to cytostatics. The effects were compared to the outcome of Bcl-xL down-regulation. Upon treatment with antisense ODNs, PKCeta and Bcl-xL were both significantly reduced on mRNA and protein level. Down-regulation of either PKCeta or Bcl-xL in combination with vincristine or paclitaxel resulted in a significant increase in caspase-3 activity compared to that in the control oligonucleotide treated cells. In addition, PKCeta down-regulation augmented vincristine-induced dissipation of mitochondrial transmembrane potential. In conclusion, these results confirm that PKCeta might represent a considerable resistance factor and an interesting target to improve anticancer chemotherapy. PMID:15159020

  11. Apoptotic Effect of Coix Polysaccharides on A549 Lung Cancer Cells in Vitro

    Luo, Cheng; Liu, Wei; Lu, Xiangyi

    2012-01-01

    Background and objective Coix seeds are commonly used in Traditional Chinese Medicine and ingested through daily diet. The aim of this study is to analyze the apoptotic effect of coix polysaccharides on A549 cells. Methods A fraction of polysaccharides was isolated from coix seeds and extracted by ethanol precipitation. The extract was then purified by dialysis and DEAE-52 ion-exchange chromatography. Cell viability was determined by the MTT assay. Cell morphology was observed by scanning ele...

  12. Apoptotic Effect of Coix Polysaccharides on A549 Lung Cancer Cells in Vitro

    Cheng LUO

    2012-11-01

    Full Text Available Background and objective Coix seeds are commonly used in Traditional Chinese Medicine and ingested through daily diet. The aim of this study is to analyze the apoptotic effect of coix polysaccharides on A549 cells. Methods A fraction of polysaccharides was isolated from coix seeds and extracted by ethanol precipitation. The extract was then purified by dialysis and DEAE-52 ion-exchange chromatography. Cell viability was determined by the MTT assay. Cell morphology was observed by scanning electronic microscopy (SEM, and cell cycle was detected by flow cytometry (FCM. The relative quantities of caspase-3 and caspase-9 were determined by RT-PCR. Results Coix polysaccharides exerted remarkable inhibitory effects on A549 cell proliferation. Apoptotic bodies were observed by SEM. Apoptotic induction was also verified by DNA accumulation using propidium iodide nucleus staining in the S phase by flow cytometry, as well as by DNA fragmentation using the comet assay. Regarding the molecular mechanism of apoptosis induction, the gene expression of caspase-3 and caspase-9 increased after coix polysaccharide treatment. Conclusion Polysaccharide fraction CP-1 induced A549 cell apoptosis.

  13. 透明质酸寡糖调节A549/DDP多药耐药作用的研究%Effects of reversing drug resistant of hyaluronate oligomers on A549/DDP cell line of human lung cancer

    张宪真; 王宝中

    2009-01-01

    Objective:To investigate the effects of hyaluronate oligomers on the multiple drug resistance of lung cancer cell lines A549/DDP. Methods: After co-culturing A549/DDP and CD44 monoclonal antibody or hyaluronan oligomers for 48 hours,to detect the following parameters:Hyaluronate contents of the medium by radioimmunoassay,MDR1,MRP,LRP expressions by flow cytometry,survival rate of cells under different concentrations of cisplatin by MTT tests. Results: Hyaluronan oligomers can decrease hyaluronan expression,and MDR1,MRP,LRP expression of A549/DDP.In addition,apoptosis level of cells treated by hyaluronan oligomers increased significantly in higher concentration cisplatin. Conclution: In vitro,hyaluronan oligomers can reverse drug resistance of A549/DDP.%目的:通过研究透明质酸寡糖对人肺腺癌细胞系A549/DDP的P糖蛋白和多药耐药相关蛋白(MRP)、肺耐药蛋白(LRP)表达的影响,探讨透明质酸在引起肿瘤细胞多药耐药中的作用.方法: 将CD44单抗或透明质酸寡糖与A549/DDP细胞共培养48小时,放免法检测培养基中细胞所分泌透明质酸的含量,流式细胞仪检测经上述处理的A549/DDP表面MDR1 、MRP、LRP的表达率,MTT法检测在不同浓度顺铂作用下,各组细胞的存活率.结果: 经透明质酸寡糖处理后A549/DDP,分泌透明质酸较未处理组明显减少(P<0.05);且细胞表面与多药耐药相关的MDR1 、MRP、LRP的表达率均降低(P<0.05).另外,处理后的细胞,在不同浓度顺铂作用时,细胞凋亡率明显增加.结论: 体外条件下,透明质酸寡糖能逆转A549/DDP的耐药.

  14. Upregulation of Id3 inhibits cell proliferation and induces apoptosis in A549/DDP human lung cancer cells in vitro.

    Chen, Fangfang; Zhao, Qinfei; Wang, Shuxia; Wang, Haiyong; Li, Xiaojun

    2016-07-01

    Inhibitor of DNA binding (Id)3 is a member of the Id multigene family of dominant‑negative helix‑loop-helix transcription factors, which function as oncogenes or tumor suppressors in human cancers. Its upregulation was recently shown to have inhibitory effects on lung cancer, which is the leading cause of cancer‑associated mortality worldwide. As drug resistance represents a major bottleneck of cancer therapy, the present study assessed the ability of Id3 to inhibit cisplatin‑resistant A549 lung adenocarcinoma cells (A549/DDP). A549/DPP cells were transiently transfected with enhanced green fluorescence protein overexpression plasmid (pEGFP) or Id3 overexpression plasmid (Id3/pEGFP), which was confirmed by confocal fluorescence microscopy, PCR and western blot analysis. The effects of Id3 on the viability and apoptosis of A549/DDP were determined using an MTT assay, fluorescence microscopy with Hoechst 33258 staining and flow cytometry following Annexin V/propidium iodide double staining. The results revealed that overexpression of Id3 significantly inhibited the proliferation and viability of A549/DDP cells in a time‑dependent manner. Furthermore, overexpression of Id3 significantly increased the apoptotic rate of A549/DDP cells from 2.73 to 16.92%, confirming the implication of Id3 in the negative control of tumour growth. The results of the present study revealed that overexpression of Id3 may serve as a novel strategy for inhibiting cisplatin‑sensitive lung cancer. Further experiments will be performed to determine whether Id3 overexpression could enhance the sensitivity of lung cancer cells to DDP. PMID:27176047

  15. Psoralidin induces autophagy through ROS generation which inhibits the proliferation of human lung cancer A549 cells

    Wenhui Hao

    2014-09-01

    Full Text Available Psoralidin (PSO, a natural furanocoumarin, is isolated from Psoralea corylifolia L. possessing anti-cancer properties. However, the mechanisms of its effects remain unclear. Herein, we investigated its anti-proliferative effect and potential approaches of action on human lung cancer A549 cells. Cell proliferation and death were measured by MTT and LDH assay respectively. Apoptosis was detected with Hoechst 33342 staining by fluorescence microscopy, Annexin V-FITC by flow cytometry and Western blot analysis for apoptosis-related proteins. The autophagy was evaluated using MDC staining, immunofluorescence assay and Western blot analyses for LC3-I and LC3-II. In addition, the reactive oxygen species (ROS generation was measured by DCFH2-DA with flow cytometry. PSO dramatically decreased the cell viabilities in dose- and time-dependent manner. However, no significant change was observed between the control group and the PSO-treated groups in Hoechst 33342 and Annexin V-FITC staining. The expression of apoptosis-related proteins was not altered significantly either. While the MDC-fluorescence intensity and the expression ratio of LC3-II/LC3-I was remarkably increased after PSO treatment. Autophagy inhibitor 3-MA blocked the production of LC3-II and reduced the cytotoxicity in response to PSO. Furthermore, PSO increased intracellular ROS level which was correlated to the elevation of LC3-II. ROS scavenger N-acetyl cysteine pretreatment not only decreased the ROS level, reduced the expression of LC3-II but also reversed PSO induced cytotoxicity. PSO inhibited the proliferation of A549 cells through autophagy but not apoptosis, which was mediated by inducing ROS production.

  16. Oxidative Stress Facilitates IFN-γ-Induced Mimic Extracellular Trap Cell Death in A549 Lung Epithelial Cancer Cells.

    Lin, Chiou-Feng; Chen, Chia-Ling; Chien, Shun-Yi; Tseng, Po-Chun; Wang, Yu-Chih; Tsai, Tsung-Ting

    2016-01-01

    We previously demonstrated that IFN-γ induces an autophagy-regulated mimic extracellular trap cell death (ETosis) in A549 human lung cancer cells. Regarding reactive oxygen species (ROS) are involved in ETosis, this study investigated the role of oxidative stress. After IFN-γ stimulation, a necrosis-like cell death mimic ETosis occurred accompanied by the inhibition of cell growth, aberrant nuclear staining, and nucleosome release. ROS were generated in a time-dependent manner with an increase in NADPH oxidase component protein expression. STAT1-mediated IFN regulatory factor-1 activation was essential for upregulating ROS production. By genetically silencing p47phox, IFN-γ-induced ROS and mimic ETosis were significantly attenuated. This mechanistic study indicated that ROS may mediate DNA damage followed by histone H3 citrullination. Furthermore, ROS promoted IFN-γ-induced mimic ETosis in cooperation with autophagy. These findings further demonstrate that ROS regulates IFN-γ-induced mimic ETosis in lung epithelial malignancy. PMID:27575372

  17. Low dose hyper-radiosensitivity in human lung cancer cell line A549 and its possible mechanisms

    Objective: To study the low dose hyper-radiosensitivity in human lung cancer cell line A549, and its possible mechanisms. Methods: Exponentially growing A549 cells were irradiated with 60Co γ-rays at doses of 0-2 Gy. Together with flow cytometry for precise cell sorting, cell survival fraction was measured by mean of conventional colony-formation assay. ATM1981 Ser-P protein expression was examined by Western blot. Apoptosis was identified by Hoechst 33258 fluorescent staining, and Annexin V-FITC and propidium iodide staining flow cytometry. Cell cycle distribution was observed by flow cytometry. Results: There was an excessive cell killing per unit dose when the doses were below about 0.3 Gy, and the cells exhibited more resistant response at the doses between 0.3 and 0.5 Gy, the cell survival fraction was decreased as the doses over 0.5 Gy. The expression of ATM1981Ser-P protein was first observed at 0.2 Gy, followed by an increase over 0.2 Gy, and reached the peak at 0.5 Gy (compared with 0.2 Gy group, t=7.96, P0.05). 24 hours after irradiation, part cells presented the characteristic morphological change of apoptosis, and the apoptosis curve was coincident with the dose-survival curve. Compared with the control group, the cell cycle had no change post-irradiation to 0.1 and 0.2 Gy. G2/M phase arrest was manifested at 6 and 12 hours post-irradiation to 0.3, 0.4 and 0.5 Gy (t=2.87, 2.88, 4.92 and 3.70, 3.12, 8.11, P2/M phase was decreased at 24 hours post-irradiation (t=3.87, 4.77, 3.01, P<0.05). Conclusions: A549 cells displays the phenomenon of hyper-radiosensitivity (HRS) /induced radioresistance (IRR). The model of cell death induced by low dose irradiation is mainly apoptosis. The activity of ATM and cell cycle change might play an important role in HRS/IRR. (authors)

  18. Low Dose Hyper-radiosensitivity in Human Lung Cancer Cell Line A549 and Its Possible Mechanisms

    Xiaofang DAI; Dan TAO; Hongge WU; Jing CHENG

    2009-01-01

    The low dose hyper-radiosensitivity (HRS) in human lung cancer cell line A549 was in-vestigated,the changes of ATM kinase,cell cycle and apoptosis of cells at different doses of radiation were observed,and the possible mechanisms were discussed.A549 cells in logarithmic growth phase were irradiated with 60Co γ-rays at doses of 0-2 Gy.Together with flow cytometry for precise cell sorting,cell survival fraction was measured by means of conventional colony-formation assay.The expression of ATM1981Ser-P protein was examined by Western blot 1 h after radiation.Apoptosis was detected by Hoechst 33258 fluorescent staining,and Annexin V-FITC/PI staining flow cytometry 24 h after radiation.Cell cycle distribution was observed by flow cytometly 6,12 and 24 h after ra-diation.The results showed that the expression of ATM1981Ser-P protein was observed at 0.2 Gy,followed by an increase at >0.2 Gy,and reached the peak at 0.5 Gy,with little further increase as the dose exceeded 0.5 Gy.Twenty-four h after radiation,partial cells presented the characteristic mor-phological changes of apoptosis,and the cell apoptosis curve was coincident with the survival curve.As compared with control group,the cell cycle almost had no changes after exposure to 0.1 and 0.2 Gy radiation (P>0.05).After exposure to 0.3,0.4 and 0.5 Cry radiation,G2/M phase arrest occurred 6 and 12 h after radiation (P<0.05),and the ratio of G2/M phase cells was decreased 24 h after radiation (P<0.05).It was concluded that A549 cells displayed the phenomenon of HRS/IRR.The mode of cell death was mainly apoptosis.The activity of ATM and cell cycle change may take an important role in HRS/IRR.

  19. 荞麦七提取物对肺癌A549细胞增殖及凋亡的影响%Effects of Fagopyrum cymosum extracts on proliferation and apoptosis of lung cancer cell line A549

    李健; 王晓梅; 杨春娟; 刘帆

    2015-01-01

    Objective To investigate the effects of Fagopyrum cymosum extracts on proliferation and apoptosis of human lung cancer cell line A549. Methods A549 lung cancer cells were processed with aqueous extracts and anthraquinone of Fagopyrum cymosum. Cell viability was detected by trypan blue staining. The inhibition rate of cell proliferation was detected by MTT. The protein expression levels of Csapase 9 and P53 were detected by immunohis-tochemical method. Results The inhibition effects of Fagopyrum cymosum aqueous extracts on lung cancer cell line A549 increased along with higher concentration of the extracts. The inhibition rate at 72 h was significantly higher than the rates at 24 h and 48 h, while there were no significant differences in inhibition rates among the three con-centrations of Fagopyrum cymosum anthraquinone. The induction on Csapase 9 and inhibition on P53 by both extracts were enhanced with the increase of concentration. Conclusion The aqueous extracts and anthraquinone of Fagopy-rum cymosum can inhibit the proliferation of human lung cancer cell line A549 and induce their apoptosis, with the underlying mechanism possibly related to the up-regulation of Caspase 9 and down-regulation of P53.%目的:研究荞麦七提取物对人肺癌A549细胞增殖及凋亡的影响。方法应用荞麦七水提物及荞麦七蒽醌处理肺癌A549细胞,锥虫蓝染色法检测细胞存活率,MTT法检测细胞增殖抑制率,免疫细胞化学法检测Caspase 9和P53蛋白表达水平。结果荞麦七水提取物对肺癌A549细胞增殖的抑制作用随浓度而增强,72 h的抑制率明显较24 h及48 h强,荞麦七蒽醌3种浓度的抑制率之间差异不大。2种提取物对Caspase 9的诱导作用均随着浓度的增大而增强,对P53的抑制作用也随着浓度的增大而增强。结论荞麦七水提物及蒽醌能抑制人肺癌A549细胞的增殖,并诱导其凋亡,其机制可能与上调Caspase 9的表达及下调P53的表达有关。

  20. Casiopeina IIgly-induced oxidative stress and mitochondrial dysfunction in human lung cancer A549 and H157 cells

    Casiopeinas are a series of mixed chelate copper complexes that are being evaluated as anticancer agents. Their effects in the cell include oxidative damage and mitochondrial dysfunction, yet the molecular mechanisms leading to such effects remain unclear. We tested whether [Cu(4,7-dimethyl-phenanthroline)(glycinate)]NO3 (Casiopeina IIgly or Cas IIgly) could alter cellular glutathione (GSH) levels by redox cycling with GSH to generate ROS and cellular oxidative stress. Cas IIgly induced a dramatic drop in intracellular levels of GSH in human lung cancer H157 and A549 cells, and is able to use GSH as source of electrons to catalyze the Fenton reaction. In both cell lines, the toxicity of Cas IIgly (2.5-5 μM) was potentiated by the GSH synthesis inhibitor L-buthionine sulfoximine (BSO) and diminished by the catalytic antioxidant manganese(III) meso-tetrakis(N,N'-diethylimidazolium-2-yl)porphyrin (MnTDE-1,3-IP5+), thus supporting an important role for oxidative stress. Cas IIgly also caused an over-production of reactive oxygen species (ROS) in the mitochondria and a depolarization of the mitochondrial membrane. Moreover, Cas IIgly produced mitochondrial DNA damage that resulted in an imbalance of the expression of the apoproteins of the mitochondrial respiratory chain, which also can contribute to increased ROS production. These results suggest that Cas IIgly initiates multiple possible sources of ROS over-production leading to mitochondrial dysfunction and cell death.

  1. Role of autophagy in the ω-3 long chain polyunsaturated fatty acid-induced death of lung cancer A549 cells

    Yao, Qinghua; Fu, Ting; Wang, Lu; LAI, YUEBIAO; Wang, Yuqi; Xu, Chao; Huang, Lulu; Guo, Yong

    2015-01-01

    The present study identified that ω-3 long chain polyunsaturated fatty acids (ω-3 PUFAs), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) demonstrate anti-proliferative effects in lung cancer A549 cells. MTS and cytotoxicity assays were conducted to confirm that ω-3 PUFAs induced cell death. Autophagy-associated gene and signaling pathways were also detected. Microtubule-associated protein light chain 3 (LC3) expression was found to be increased subsequent to treatment with DHA and...

  2. The Study on Anti-cancer Effects of Distilling Fresh-ginseng Herbal acupuncture against implanted Sarcoma-180 in vivo and A549 human epithelial lung cancer cells in vitro

    Hae-Young Jang; Ki-Rok Kwon; Hee-Soo Park

    2004-01-01

    Objectives : This study was to investigate the anti-cancer effects of herbal acupuncture with distilled fresh ginseng. The herbal acupuncture was injected to Chung-wan(C.V12) and Wisu(BL21) of mice that were subjected to Sarcoma-180 abdominal cancer cell and A549 human epithelial lung cancer cells in vitro. Methods : Anti-cancer effects of distilled fresh ginseng herbal acupuncture were tested by measruing Cox, Bcl-2, and Bax by using RT-PCR in A549 human epithelial lung cancer cells in v...

  3. Induction of Apoptotic Effects of Antiproliferative Protein from the Seeds of Borreria hispida on Lung Cancer (A549 and Cervical Cancer (HeLa Cell Lines

    S. Rupachandra

    2014-01-01

    Full Text Available A 35 KDa protein referred to as F3 was purified from the seeds of Borreria hispida by precipitation with 80% ammonium sulphate and gel filtration on Sephadex G-100 column. RP-HPLC analysis of protein fraction (F3 on an analytical C-18 column produced a single peak, detected at 220 nm. F3 showed an apparent molecular weight of 35 KDa by SDS PAGE and MALDI-TOF-MS analyses. Peptide mass fingerprinting analysis of F3 showed the closest homology with the sequence of 1-aminocyclopropane-1-carboxylate deaminase of Pyrococcus horikoshii. The protein (F3 exhibited significant cytotoxic activity against lung (A549 and cervical (HeLa cancer cells in a dose-dependent manner at concentrations ranging from 10 µg to 1000 µg/mL, as revealed by the MTT assay. Cell cycle analysis revealed the increased growth of sub-G0 population in both cell lines exposed to a concentration of 1000 µg/mL of protein fraction F3 as examined from flow cytometry. This is the first report of a protein from the seeds of Borreria hispida with antiproliferative and apoptotic activity in lung (A549 and cervical (HeLa cancer cells.

  4. Investigation of radiation-induced transcriptome profile of radioresistant non-small cell lung cancer A549 cells using RNA-seq.

    Hee Jung Yang

    Full Text Available Radioresistance is a main impediment to effective radiotherapy for non-small cell lung cancer (NSCLC. Despite several experimental and clinical studies of resistance to radiation, the precise mechanism of radioresistance in NSCLC cells and tissues still remains unclear. This result could be explained by limitation of previous researches such as a partial understanding of the cellular radioresistance mechanism at a single molecule level. In this study, we aimed to investigate extensive radiation responses in radioresistant NSCLC cells and to identify radioresistance-associating factors. For the first time, using RNA-seq, a massive sequencing-based approach, we examined whole-transcriptome alteration in radioresistant NSCLC A549 cells under irradiation, and verified significant radiation-altered genes and their chromosome distribution patterns. Also, bioinformatic approaches (GO analysis and IPA were performed to characterize the radiation responses in radioresistant A549 cells. We found that epithelial-mesenchymal transition (EMT, migration and inflammatory processes could be meaningfully related to regulation of radiation responses in radioresistant A549 cells. Based on the results of bioinformatic analysis for the radiation-induced transcriptome alteration, we selected seven significant radiation-altered genes (SESN2, FN1, TRAF4, CDKN1A, COX-2, DDB2 and FDXR and then compared radiation effects in two types of NSCLC cells with different radiosensitivity (radioresistant A549 cells and radiosensitive NCI-H460 cells. Interestingly, under irradiation, COX-2 showed the most significant difference in mRNA and protein expression between A549 and NCI-H460 cells. IR-induced increase of COX-2 expression was appeared only in radioresistant A549 cells. Collectively, we suggest that COX-2 (also known as prostaglandin-endoperoxide synthase 2 (PTGS2 could have possibility as a putative biomarker for radioresistance in NSCLC cells.

  5. 1‑O‑acetylbritannilactone combined with gemcitabine elicits growth inhibition and apoptosis in A549 human non‑small cell lung cancer cells.

    Wang, Feng; Li, Hong; Qiao, Jian-Ou

    2015-10-01

    Non‑small‑cell lung cancer (NSCLC) accounts for ~85% of all lung cancer cases, with a 5‑year survival rate of britannica, a Chinese traditional medicine, has been demonstrated to have anticancer activity. In the present study, the antiproliferative and proapoptotic abilities of ABL alone or in combination with gemcitabine in a human NSCLC cell line were investigated. A549 cells were treated in vitro with ABL, gemcitabine, and a combination of ABL and gemcitabine for 72 h. The results demonstrated that ABL and gemcitabine inhibited cell growth and induced apoptosis of A549 cells. These effects were more potent following the combination of ABL and gemcitabine treatment than either agent alone. Furthermore, the signal transduction analysis revealed nuclear factor (NF)‑κB expression was significantly decreased by ABL and the combination treatment. The inhibitor nuclear factor κBα (IκBα) and Bax levels were upregulated whereas Bcl‑2 was substantially downregulated following treatment. The present findings suggest that ABL combined with gemcitabine elicits potent apoptosis of lung cancer cells and therefore, ABL has the potential to be developed as a chemotherapeutic agent. PMID:26151622

  6. Quercetin Down-regulates IL-6/STAT-3 Signals to Induce Mitochondrial-mediated Apoptosis in a Non-small-cell Lung-cancer Cell Line, A549

    Avinaba Mukherjee

    2015-03-01

    Full Text Available Objectives: Quercetin, a flavonoid compound, has been reported to induce apoptosis in cancer cells, but its anti-inflammatory effects, which are also closely linked with apoptosis, if any, on non-small-cell lung cancer (NSCLC have not so far been critically examined. In this study, we tried to determine if quercetin had any demonstrable anti-inflammatory potential, which also could significantly contribute to inducing apoptosis in a NSCLC cell line, A549. Methods: In this context, several assays, including cytotoxicity, flow cytometry and fluorimetry, were done. Gene expression was analyzed by using a western blot analysis. Results: Results revealed that quercetin could induce apoptosis in A549 cells through mitochondrial depolarization by causing an imbalance in B-cell lymphoma 2/Bcl2 Antagonist X (Bcl2/Bax ratio and by down-regulating the interleukine-6/signal transducer and activator of transcription 3 (IL-6/STAT3 signaling pathway. An analysis of the data revealed that quercetin could block nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB activity at early hours, which might cause a down-regulation of the IL-6 titer, and the IL-6 expression, in turn, could inhibit p-STAT3 expression. Down-regulation of both the STAT3 and the NF-κB expressions might, therefore, cause down-regulation of Bcl2 activity because both are major upstream effectors of Bcl2. Alteration in Bcl2 responses might result in an imbalance in the Bcl2/Bax ratio, which could ultimately bring about mitochondria mediated apoptosis in A549 cells. Conclusion: Overall, the finding of this study indicates that a quercetin induced anti-inflammatory pathway in A549 cells appeared to make a significant contribution towards induction of apoptosis in NSCLC and, thus, may have a therapeutic use such as a strong apoptosis inducer in cancer cells.

  7. Biological effects of heavy ion and X-ray irradiation on lung cancer cells A549%重离子与X射线照射肺癌细胞A549的生物学效应比较

    杨立娜; 冉俊涛; 张红; 刘圆圆; 孙超; 张秋宁; 王新宇; 王小虎

    2014-01-01

    Objective To compare the effects of carbon heavy ion and X-ray irradiation on survival fraction,cell cycle,cell apoptosis and expression of DNA-PKcs of A549 lung cancer cells.Methods A549 cells were irradiated by carbon heavy ion and X-ray.Cell survival fraction,cell cycle and apoptosis were analyzed by clonogenic formation assay,flow cytometry and Hoechst 33258 staining,respectively.Real time-PCR was performed to detect the expressions of DNA-PKcs and H2AX mRNA.Results Lower cell survival fraction,more G2/M phase arrest and higher apoptosis rate were detected in the A549 cells exposed to carbon heavy ion than X-ray(t =4.77,14.53,14.54,P < 0.05).Expression of DNA-PKcs was up-regulated after irradiation to carbon heavy ion and X-ray(t =10.91,5.05,P < 0.05).Conclusions Both heavy ion and X-ray irradiations enhance the expression of DNA-PKcs,induce apoptosis through regulating cell cycle arrest,and hence reduce survival of A549 cells.Heavy ion irradiation shows more stronger biological effects than X-ray irradiation.%目的 比较碳重离子与X射线对肺癌细胞的生物学效应.方法 对A549细胞分别进行碳重离子和X射线照射,通过克隆形成实验检测照射后细胞存活情况;流式细胞术检测细胞周期分布;通过Hoechst 33258荧光染料对照射后固定的细胞进行染色,计算凋亡率;采用实时荧光定量PCR方法检测照射后48 h细胞内DNA依赖性蛋白激酶催化亚单位(DNA-PKcs)和H2AX的mRNA表达水平.结果 细胞存活曲线显示,碳重离子造成的细胞存活分数远低于X射线,并将细胞周期阻滞于G2/M期(t=4.77、14.53、14.54,P<0.05),导致大部分细胞进入凋亡途径.碳重离子与X射线辐照后DNA-PKcs的表达上调(t=10.91、5.05,P<0.05).结论 碳重离子照射对肺癌细胞造成生物学效应远高于X射线.

  8. Reversal of multidrug resistance by cisplatin-loaded magnetic Fe3O4 nanoparticles in A549/DDP lung cancer cells in vitro and in vivo

    Li K

    2013-05-01

    Full Text Available Ke Li,1 Baoan Chen,1,2 Lin Xu,3 Jifeng Feng,3 Guohua Xia,1,2 Jian Cheng,1,2 Jun Wang,1,2 Feng Gao,1,2 Xuemei Wang,41Department of Hematology, Key Medical Disciplines of Jiangsu Province, Zhongda Hospital, Medical School, Southeast University, Nanjing, 2Faculty of Oncology, Medical School, Southeast University, Nanjing, 3Department of Thoracic Surgery, Jiangsu Province Cancer Hospital, Jiangsu Province, 4State Key Laboratory of Bioelectronics, Southeast University, Nanjing, People’s Republic of ChinaAbstract: The purpose of this study was to explore whether magnetic Fe3O4 nanoparticles (Fe3O4-MNP loaded with cisplatin (Fe3O4-MNP-DDP can reverse DDP resistance in lung cancer cells and to investigate mechanisms of multidrug resistance in vitro and in vivo. MTT assay showed that DDP inhibited both A549 cells and DDP-resistant A549 cells in a time-dependent and dose-dependent manner, and that this inhibition was enhanced by Fe3O4-MNP. An increased rate of apoptosis was detected in the Fe3O4-MNP-DDP group compared with a control group and the Fe3O4-MNP group by flow cytometry, and typical morphologic features of apoptosis were confirmed by confocal microscopy. Accumulation of intracellular DDP in the Fe3O4-MNP-DDP group was greater than that in the DDP group by inductively coupled plasma mass spectrometry. Further, lower levels of multidrug resistance-associated protein-1, lung resistance-related protein, Akt, and Bad, and higher levels of caspase-3 genes and proteins, were demonstrated by reverse transcriptase polymerase chain reaction and Western blotting in the presence of Fe3O4-MNP-DDP. We also demonstrated that Fe3O4-MNP enhanced the effect of DDP on tumor growth in BALB/c nude mice bearing DDP-resistant human A549 xenografts by decreasing localization of lung resistance-related protein and Ki-67 immunoreactivity in cells. There were no apparent signs of toxicity in the animals. Overall, these findings suggest potential clinical application of

  9. Ameliorative effects of dimetylthiourea and N-acetylcysteine on nanoparticles induced cyto-genotoxicity in human lung cancer cells-A549.

    Ritesh Kumar Srivastava

    Full Text Available We study the ameliorative potential of dimetylthiourea (DMTU, an OH• radical trapper and N-acetylcysteine (NAC, a glutathione precursor/H₂O₂ scavenger against titanium dioxide nanoparticles (TiO₂-NPs and multi-walled carbon nanotubes (MWCNTs induced cyto-genotoxicity in cultured human lung cancer cells-A549. Cytogenotoxicity was induced by exposing the cells to selected concentrations (10 and 50 µg/ml of either of TiO₂-NPs or MWCNTs for 24 h. Anti-cytogenotoxicity effects of DMTU and NAC were studied in two groups, i.e., treatment of 30 minutes prior to toxic insult (short term exposure, while the other group received DMTU and NAC treatment during nanoparticles exposure, i.e., 24 h (long term exposure. Investigations were carried out for cell viability, generation of reactive oxygen species (ROS, micronuclei (MN, and expression of markers of oxidative stress (HSP27, CYP2E1, genotoxicity (P⁵³ and CYP2E1 dependent n- nitrosodimethylamine-demethylase (NDMA-d activity. In general, the treatment of both DMTU and NAC was found to be effective significantly against TiO₂-NPs and MWCNTs induced cytogenotoxicity in A549 cells. Long-term treatment of DMTU and NAC during toxic insults has shown better prevention than short-term pretreatment. Although, cells responded significantly to both DMTU and NAC, but responses were chemical specific. In part, TiO₂-NPs induced toxic responses were mediated through OH• radicals generation and reduction in the antioxidant defense system. While in the case of MWCNTs, adverse effects were primarily due to altering/hampering the enzymatic antioxidant system. Data indicate the applicability of human lung cancer cells-A549 as a pre-screening tool to identify the target specific prophylactic and therapeutic potential of drugs candidate molecules against nanoparticles induced cellular damages.

  10. The Effects of Davallic Acid from Davallia divaricata Blume on Apoptosis Induction in A549 Lung Cancer Cells

    Tsu-Liang Chang; Kai-Yu Chen; Kai-Hsien Chen; Yu-Hsiang Cheng; We-Chang Chang; An-Sheng Cheng

    2012-01-01

    Traditional or folk medicinal herbs continue to be prescribed in the treatment of various diseases and conditions in many cultures. Recent scientific efforts have focused on the potential roles of extracts of traditional herbs as alternative and complementary medications for cancer treatment. In Taiwan, Davallia divaricata Blume has been traditionally employed in folk medicine for therapy of lung cancer, davallic acid being the major active compound of D. divaricata Blume. In this study, we i...

  11. Encapsulated paclitaxel nanoparticles exhibit enhanced anti-tumor efficacy in A549 non-small lung cancer cells.

    Huang, Guojin; Zang, Bao; Wang, Xiaowei; Liu, Gang; Zhao, Jianqiang

    2015-12-01

    In the present study, paclitaxel (PTX) were encapsulated with polyethylene glycol (PEG)-polylactide (PLA)/D-α tocopheryl polyethylene glycol 1000 succinate (TPGS) (PEG-PLA/TPGS) and the enhanced anti-tumor activity of this PTX mixed micelles (PTX-MM) was evaluated in lung cancer cells. The PTX-MM prepared by a solvent evaporation method was demonstrated to have high drug-loading efficiency (23.2%), high encapsulation efficiency (76.4%), and small size (59 nm). In vitro release assay showed the slow release behavior of PTX-MM, suggesting the good stability of the PTX-MM essential for long circulation time. In vitro kinetics assay demonstrated that PTX-MM could promote absorption and increase relative bioavailability. The anti-cancer efficiency of PTX-MM was also examined by both in vitro and in vivo studies. PTX-MM exhibits obvious cytotoxicity against lung cancer cells with much lower IC50 value when compared with commercial formulated PTX or PTX + TPGS. The xenograft tumor model studies on nude mice indicated that PTX-MM inhibits tumor growth more effectively than other formulations. It was also found that most of mixed micelles were integral in tumor site to exhibit anti-cancer activity. Our results suggested that the use of PTX-MM as an anti-cancer drug may be an effective approach to treat lung cancer. PMID:26525950

  12. MiR-92b regulates the cell growth, cisplatin chemosensitivity of A549 non small cell lung cancer cell line and target PTEN.

    Li, Yan; Li, Li; Guan, Yan; Liu, Xiuju; Meng, Qingyong; Guo, Qisen

    2013-11-01

    MicroRNAs (miRNAs) have emerged to play important roles in tumorigenesis and drug resistance of human cancer. Fewer studies were explored the roles of miR-92b on human lung cancer cell growth and resistance to cisplatin (CDDP). In this paper, we utilized real-time PCR to verify miR-92b was significantly up-regulated in non-small cell lung cancer (NSCLC) tissues compared to matched adjacent normal tissues. In vitro assay demonstrated that knock-down of miR-92b inhabits cell growth and sensitized the A549/CDDP cells to CDDP. Furthermore, we found miR-92b could directly target PTEN, a unique tumor suppressor gene, which was downregulated in lung cancer tissues compared to the matched adjacent normal tissues. These data indicate that the miR-92b play an oncogene roles by regulates cell growth, cisplatin chemosensitivity phenotype, and could serve as a novel potential maker for NSCLC therapy. PMID:24099768

  13. Impact of CHK2-small interfering RNA on CpG ODN7909-enhanced radiosensitivity in lung cancer A549 cells

    Chen W

    2012-12-01

    Full Text Available Wei Chen,* Xiaoqun Liu,* Tiankui Qiao, Sujuan Yuan Department of Oncology, Jinshan Hospital, Fudan University, Shanghai, People's Republic of China*These authors contributed equally to this workObjective: To investigate the impact of checkpoint kinase 2 (CHK2-small interfering RNA (CHK2-siRNA on the enhancement of radiosensitivity by CpG oligodeoxynucleotide (ODN 7909 in lung cancer A549 cells.Methods: The A549 cells were randomly divided into five groups: control, CpG, X-ray, CpG + X-ray, and CHK2-siRNA + CpG + X-ray. Cell colonization was observed using inverted microscopy. Cell cycle and apoptosis were analyzed by flow cytometry. CHK2 expression was detected by Western blot. CHK2-siRNA was adopted to silence the expression of CHK2.Results: The level of CHK2 phosphorylation was higher in the CpG + X-ray group than in the X-ray group. Increases in G2/mitotic (M phase arrest and apoptosis and a decrease of cell survival rate in the CpG + X-ray group were statistically significant (P < 0.05 when compared with the CHK2-siRNA + CpG + X-ray group in which the expression of CHK2 was obviously inhibited. The combination of CpG ODN7909 and X-ray irradiation was found to enhance the mitotic death of A549 cells. The sensitization enhancement ratio of mean death dose (D0 was 1.42 in the CpG + X-ray group, which was higher than that of the CHK2-siRNA + CpG + X-ray group, in which D0 was 1.05.Conclusion: To a certain extent, the impact of a combination of CpG ODN7909 and X-ray on G2/M phase arrest, apoptosis, and rate of cell survival was attenuated by CHK2-siRNA in human lung adenocarcinoma A549 cells, indicating that increased phosphorylation of CHK2 might be a radiosensitive pathway.Keywords: oligodeoxynucleotide, checkpoint kinase 2, mitotic death, apoptosis, X-ray

  14. Andrographolide down-regulates hypoxia-inducible factor-1α in human non-small cell lung cancer A549 cells

    Andrographolide (Andro), a diterpenoid lactone isolated from a traditional herbal medicine Andrographis paniculata, is known to possess multiple pharmacological activities. In our previous study, Andro had been shown to inhibit non-small cell lung cancer (NSCLC) A549 cell migration and invasion via down-regulation of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Here we demonstrated that Andro inhibited the expression of hypoxia-inducible factor-1α (HIF-1α) in A549 cells. HIF-1α plays an important role in tumor growth, angiogenesis and lymph node metastasis of NSCLC. The Andro-induced decrease of cellular protein level of HIF-1α was correlated with a rapid ubiquitin-dependent degradation of HIF-1α, and was accompanied by increased expressions of hydroxyl-HIF-1α and prolyl hydroxylase (PHD2), and a later decrease of vascular endothelial growth factor (VEGF) upon the treatment of Andro. The Andro-inhibited VEGF expression appeared to be a consequence of HIF-1α inactivation, because its DNA binding activity was suppressed by Andro. Molecular data showed that all these effects of Andro might be mediated via TGFβ1/PHD2/HIF-1α pathway, as demonstrated by the transfection of TGFβ1 overexpression vector and PHD2 siRNA, and the usage of a pharmacological MG132 inhibitor. Furthermore, we elucidated the involvement of Andro in HIF-1α transduced VEGF expression in A549 cells and other NSCLC cell lines. In conclusion, these results highlighted the potential effects of Andro, which may be developed as a chemotherapeutic or an anti-angiogenesis agent for NSCLC in the future.

  15. Aurora A反义寡核苷酸对肺癌细胞A549的作用和对紫杉醇化疗敏感性的影响%The effect of antisense oligodeoxynucleotides targeting Aurora A kinase on cell proliferation and chemosensitivity to paclitaxel in human lung cancer cell line A549

    Rui Meng; Gang Wu; Jing Cheng; Tao Wang

    2007-01-01

    Objective:Aurora A kinase representing a family of evolutionarily conserved mitotic serine/threonine kinases has been found elevated in human lung adenocarcinoma cell line A549.It is suggested that the overexpression of Aurora A contributes to the carcinogenesis,chromosomal instability (CIN),and de-differentiation of lung cancers.To address its possibility as a therapeutic target for lung cancer,we employed the antisense oligodeoxynucleotide (ASODN) technique to inhibi Aurora A expression and investigate its effects on tumor growth and cell cycle of A549.as well as the chemosensitivilty to paclitaxel.Methods:Aurora A ASODN was synthesized and transfected into A549 cells by lipofectAMINE 2000.Aurora A mRNA and protein expression were examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively.Cell cycle distribution was observed by flow cytometer.MTT assay was used to evaluate cell inhibition ratio before and after transfection.Results:The proliferation of the A549 cell swas inhibited by Aurora AASODN dose and time dependently.It was also observed thal the IC50 of A549 cells after 48 hours'treatmenl of ASODN was about 300 nmol/L and under such circumstances,the Aurora A mRNA and protein expression significantly decreased(P<0.05),along with the induction of accumulation of cells in S phase and the G2-M transition.Furlhermore.cell inhibition ratio of the combination of Aurora AASODN and paclitaxel was higher significantly than paclitaxel(P<0.05)or Aurora AASODN alone (P<0.05).Conclusion:Inhibition of Aurora A expression can result in the suppression of cell growth and chemosensilizina activity to paclitaxel in human lung cancer cell line A549.

  16. Quercetin metabolites inhibit MMP-2 expression in A549 lung cancer cells by PPAR-γ associated mechanisms.

    Chuang, Cheng-Hung; Yeh, Chiao-Lin; Yeh, Shu-Lan; Lin, En-Shyh; Wang, Li-Yu; Wang, Ying-Hsuna

    2016-07-01

    Our previous study demonstrated that quercetin-metabolite-enriched plasma (QP) but not quercetin itself upregulates peroxisome proliferator-activated receptor gamma (PPAR-γ) expression to induce G2/M arrest in A549 cells. In the present study, we incubated A549 cells with QP as well as quercetin-3-glucuronide (Q3G) and quercetin-3'-sulfate (Q3'S), two major metabolites of quercetin, to investigate the effects of quercetin metabolites on cell invasion and migration, the possible mechanisms and the role of PPAR-γ. We also compared the effects of QP with those of quercetin and troglitazone (TGZ), a PPAR-γ ligand. The results showed that QP significantly suppressed cell invasion and migration, as well as matrix metalloproteinases (MMPs)-2 activity and expression in a dose-dependent manner. The effects of 10% QP on those parameters were similar to those of 10μM quercetin and 20μM TGZ. However, QP and TGZ rather than quercetin itself increased the expressions of nm23-H1 and tissue inhibitor of metalloproteinase (TIMP-2). Furthermore, we demonstrated that Q3G and Q3'S also inhibited the protein expression of MMP-2. GW9662, a PPAR-γ antagonist, significantly diminished such an effect of Q3G and Q3'S. Silencing PPAR-γ expression in A549 cells also significantly diminished the suppression effect of Q3G and Q3'S on MMP-2 expression. Taken together, our study demonstrated that QP inhibited cell invasion and migration through nm23-H1/TIMP-2/MMP-2 associated mechanisms. The upregulation of PPAR-γ by quercetin metabolites such as Q3G and Q3'S could play an important role in the effects of QP. PMID:27260467

  17. 肺癌A549放射抗拒细胞亚系的建立及抗拒机制的研究%Establishment of a radioresistant human lung cancer cell subline and its mechanism of radioresistance

    赵伟; 王琼; 刘莉; 石星; 丁乾; 伍钢

    2008-01-01

    Objective To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays:A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones,together with its parental A549 cells were measured by clone formation assay and flow cytometry.The mRNA and protein levels of Notch1 in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G0/G1 phase (P<0.05). The expression of Notch1 in A549-S1 was enhanced obviously than in A549 cells. But for A549-S2 the radioseasitivity was slightly increased compared with the parental cells with D0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05) .The expression of Notch1 had no marked change compared to A549 cell. Conclusions The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlation with the expression of Notch1.%目的 建立肺癌细胞系A549的放射抗拒模型并探

  18. Overexpression of the lung cancer-prognostic miR-146b microRNAs has a minimal and negative effect on the malignant phenotype of A549 lung cancer cells.

    Santosh Kumar Patnaik

    Full Text Available INTRODUCTION: Expression levels of miR-146b-5p and -3p microRNAs in human non-small cell lung cancer (NSCLC are associated with recurrence of the disease after surgery. To understand this, the effect of miR-146b overexpression was studied in A549 human lung cancer cells. METHODS: A549 cells, engineered with lentiviruses to overexpress the human pre-miR-146b precursor microRNA, were examined for proliferation, colony formation on plastic surface and in soft agar, migration and invasiveness in cell culture and in vivo in mice, chemosensitivity to cisplatin and doxorubicin, and global gene expression. miR-146b expressions were assessed in microdissected stroma and epithelia of human NSCLC tumors. Association of miR-146b-5p and -3p expression in early stage NSCLC with recurrence was analyzed. PRINCIPAL FINDINGS: A549 pre-miR-146b-overexpressors had 3-8-fold higher levels of both miR-146b microRNAs than control cells. Overexpression did not alter cellular proliferation, chemosensitivity, migration, or invasiveness; affected only 0.3% of the mRNA transcriptome; and, reduced the ability to form colonies in vitro by 25%. In human NSCLC tumors, expression of both miR-146b microRNAs was 7-10-fold higher in stroma than in cancerous epithelia, and higher miR-146b-5p but lower -3p levels were predictive of recurrence. CONCLUSIONS: Only a minimal effect of pre-miR-146b overexpression on the malignant phenotype was seen in A549 cells. This could be because of opposing effects of miR-146b-5p and -3p overexpression as suggested by the conflicting recurrence-predictive values of the two microRNAs, or because miR-146b expression changes in non-cancerous stroma and not cancerous epithelia of tumors are responsible for the prognostic value of miR-146b.

  19. Mimulone-induced autophagy through p53-mediated AMPK/mTOR pathway increases caspase-mediated apoptotic cell death in A549 human lung cancer cells.

    Hyun-Kyu An

    Full Text Available Anticancer properties and mechanisms of mimulone (MML, C-geranylflavonoid isolated from the Paulownia tomentosa fruits, were firstly elucidated in this study. MML prevented cell proliferation in a dose- and time-dependent way and triggered apoptosis through the extrinsic pathway in A549 human lung adenocarcinoma cells. Furthermore, MML-treated cells displayed autophagic features, such as the formation of autophagic vacuoles, a primary morphological feature of autophagy, and the accumulation of microtubule-associated protein 1 light chain 3 (LC3 puncta, another typical maker of autophagy, as determined by FITC-conjugated immunostaining and monodansylcadaverine (MDC staining, respectively. The expression levels of LC3-I and LC3-II, specific markers of autophagy, were also augmented by MML treatment. Autophagy inhibition by 3-methyladenine (3-MA, pharmacological autophagy inhibitor, and shRNA knockdown of Beclin-1 reduced apoptotic cell death induced by MML. Autophagic flux was not significantly affected by MML treatment and lysosomal inhibitor, chloroquine (CQ suppressed MML-induced autophagy and apoptosis. MML-induced autophagy was promoted by decreases in p53 and p-mTOR levels and increase of p-AMPK. Moreover, inhibition of p53 transactivation by pifithrin-α (PFT-α and knockdown of p53 enhanced induction of autophagy and finally promoted apoptotic cell death. Overall, the results demonstrate that autophagy contributes to the cytotoxicity of MML in cancer cells harboring wild-type p53. This study strongly suggests that MML is a potential candidate for an anticancer agent targeting both autophagy and apoptotic cell death in human lung cancer. Moreover, co-treatment of MML and p53 inhibitor would be more effective in human lung cancer therapy.

  20. Mimulone-induced autophagy through p53-mediated AMPK/mTOR pathway increases caspase-mediated apoptotic cell death in A549 human lung cancer cells.

    An, Hyun-Kyu; Kim, Kyoung-Sook; Lee, Ji-Won; Park, Mi-Hyun; Moon, Hyung-In; Park, Shin-Ji; Baik, Ji-Sue; Kim, Cheorl-Ho; Lee, Young-Choon

    2014-01-01

    Anticancer properties and mechanisms of mimulone (MML), C-geranylflavonoid isolated from the Paulownia tomentosa fruits, were firstly elucidated in this study. MML prevented cell proliferation in a dose- and time-dependent way and triggered apoptosis through the extrinsic pathway in A549 human lung adenocarcinoma cells. Furthermore, MML-treated cells displayed autophagic features, such as the formation of autophagic vacuoles, a primary morphological feature of autophagy, and the accumulation of microtubule-associated protein 1 light chain 3 (LC3) puncta, another typical maker of autophagy, as determined by FITC-conjugated immunostaining and monodansylcadaverine (MDC) staining, respectively. The expression levels of LC3-I and LC3-II, specific markers of autophagy, were also augmented by MML treatment. Autophagy inhibition by 3-methyladenine (3-MA), pharmacological autophagy inhibitor, and shRNA knockdown of Beclin-1 reduced apoptotic cell death induced by MML. Autophagic flux was not significantly affected by MML treatment and lysosomal inhibitor, chloroquine (CQ) suppressed MML-induced autophagy and apoptosis. MML-induced autophagy was promoted by decreases in p53 and p-mTOR levels and increase of p-AMPK. Moreover, inhibition of p53 transactivation by pifithrin-α (PFT-α) and knockdown of p53 enhanced induction of autophagy and finally promoted apoptotic cell death. Overall, the results demonstrate that autophagy contributes to the cytotoxicity of MML in cancer cells harboring wild-type p53. This study strongly suggests that MML is a potential candidate for an anticancer agent targeting both autophagy and apoptotic cell death in human lung cancer. Moreover, co-treatment of MML and p53 inhibitor would be more effective in human lung cancer therapy. PMID:25490748

  1. 多肽修饰载紫杉醇脂质体靶向A549肺癌干细胞的研究%Study on the ability of specific-binding peptide modified liposome loaded paclitaxel targeting A549 lung cancer stem cell

    蔡华荣; 江跃全

    2014-01-01

    Objective To prepare CD133 specific-binding peptide conjugated liposome loaded paclitaxel and evaluate the efficiency of cellular uptake and the ability of inhibiting A549 lung cancer stem cell.Methods Liposomes were prepared by film-ultrasonic method.The partical size,zeta-potential and entrapment efficiency of liposomes were evaluated.Cellular uptake effciency of A549 lung cancer stem cell for liposomes were explored.The anti-proliferation efficiency of TLP-PTX to A549 lung cancer stem cell was evaluated by MTT assay.Tumor spheroids were used to evaluate anti-tumor ability of TLP-PTX to A549 lung cancer stem cell. Results The particle diameter of TLP-PTX was (115.8 ±8.3)nm and the entrapment efficiency of PTX was 88.5%.CD133 specific-binding peptide could enhance the efficiency of cellar uptake.The uptaken efficiency of TLP by A549 lung cancer stem cell were 2.6 times higher than that of LP(P<0.05 ).The MTT Results showed that the toxicity of TLP-PTX on A549 lung cancer stem cell was significantly stronger than LP-PTX and paclitaxel solution(P<0.05 ).The tumor inhibition test results showed that TLP-PTX has good anti-tumor effect. Conclusion TLP-PTX can specifically recognize the surface marker CD133 of A549 lung cancer stem cell,facilitate liposomes into cells and inhibit A549 lung cancer stem cell proliferation.TLP-PTX is an effective drug delivery system targeting to A549 lung cancer stem cell.%目的:制备与肺癌干细胞标志物CD133具有高度亲和力的多肽修饰载紫杉醇脂质体(CD133 specific-binding peptide conjugated paclitaxel loaded liposome,TLP-PTX),考察TLP-PTX与A549肺癌干细胞的结合能力及其对A549肺癌干细胞和肺癌干细胞移植瘤的抑制作用。方法采用薄膜分散法制备TLP-PTX,观察其粒径,电位及紫杉醇的包封率等理化性质。采用细胞摄取实验和肿瘤球穿透实验考察TLP-PTX与A549肺癌干细胞的亲和力。通过MTT实验和肺癌干细胞肿瘤球抑制实

  2. Dihydroartemisinin inhibits cell proliferation via AKT/GSK3β/cyclinD1 pathway and induces apoptosis in A549 lung cancer cells

    Liao, Kui; Li, Juan; Wang, Zhiling

    2014-01-01

    Lung cancer is the most common cause of cancer-related death in the world. The main types of lung cancer are small cell lung carcinoma (SCLC) and non-small-cell lung carcinoma (NSCLC); non small cell lung carcinoma (NSCLC) includes squamous cell carcinoma (SCC), adenocarcinoma and large cell carcinoma, Non small cell lung carcinoma accounts for about 80% of the total lung cancer cases. Dihydroartemisinin (DHA) inhibits cell proliferation and induces apoptosis in several cancer cell lines. The...

  3. Empirical study on the anti-proliferation effect of siRNA against pokemon on human lung cancer cell line A549%siRNA干扰Pokemon基因影响A549细胞增殖的实验研究

    谢勇; 江涛

    2012-01-01

    目的 研究siRNA干扰Pokemon基因对肺腺癌A549细胞增殖抑制效应的变化.方法 专业设计合成3条针对Pokemon的siRNA,分别转染A549细胞后,RT-PCR检测转录水平Pokemon mRNA表达的变化,筛选出其中最高效的1条siRNA;用MTT法检测该siRNA干扰Pokemon对A549细胞增殖的抑制作用;流式细胞技术检测该siRNA干扰Pokemon对A549细胞凋亡的影响.结果 3条siRNA均成功转染A549细胞,倒置荧光显微镜下观察细胞呈圆绿色.RT-PCR结果显示有2条siRNA使细胞中Pokemon的mRNA表达降低(P<0.05).MTT法结果显示siRNA干扰Pokemon后对A549细胞增殖有抑制作用(P<0.05),其中48 h抑制效率达(24.14±1.39)%.流式细胞技术检测结果显示该siRNA干扰Pokemon可增加A549细胞的凋亡,凋亡率为14.05%.结论 应用RNA干扰Pokemon基因能够抑制A549细胞的增殖,促进A549细胞的凋亡.Pokemon基因有可能成为肺癌治疗中的一个新靶点.%[Objective] To investigate the anti-proliferation effect of siRNA against pokemon on human lung cancer cell line AS49. [Methods] We professionally devised and synthesized three siRNAs against pokemon, then transfected it into A549 cells. We detected the mRNA expression of transcription level changes by RT-PCR experiment and screened out the most efficient siRNA. The effects on cell proliferation and apoptosis were analyzed by MTT assay and flow cytometry. [Results] The three siRNAs were successfully transfected into A549 cells and the cells were green under fluorescence microscope cylindrical. The experiment of RT-PCR showed that the expression of pokemon mRNA in two siRNA groups were lower than the control groups. The MTT assay showed that the anti-proliferation effect of siRNA on A549 cells were significant (P<0.05), the anti-proliferation rate at 48 h were (24.14±1.39)%. Flow cytometry revealed that the siRNA on the A549 cells increased the apoptotic rate, the rate was 14.05%. [Conclusions] Application of RNA

  4. The repair capacity of lung cancer cell lines A549 and H1299 depends on HMGB1 expression level and the p53 status.

    Yusein-Myashkova, Shazie; Stoykov, Ivan; Gospodinov, Anastas; Ugrinova, Iva; Pasheva, Evdokia

    2016-07-01

    Elucidation of the cellular components responsive to chemotherapeutic agents as cisplatin rationalizes the strategy for anticancer chemotherapy. The removal of the cisplatin/DNA lesions gives the chance to the cancer cells to survive and compromises the chemotherapeutical treatment. Therefore, the cell repair efficiency is substantial for the clinical outcome. High mobility group box 1 (HMGB1) protein is considered to be involved in the removal of the lesions as it binds with high affinity to cisplatin/DNA adducts. We demonstrated that overexpression of HMGB1 protein inhibited cis-platinated DNA repair in vivo and the effect strongly depended on its C-terminus. We registered increased levels of DNA repair after HMGB1 silencing only in p53 defective H1299 lung cancer cells. Next, introduction of functional p53 resulted in DNA repair inhibition. H1299 cells overexpressing HMGB1 were significantly sensitized to treatment with cisplatin demonstrating the close relation between the role of HMGB1 in repair of cis-platinated DNA and the efficiency of the anticancer drug, the process being modulated by the C-terminus. In A549 cells with functional p53, the repair of cisplatin/DNA adducts is determined by а complex action of HMGB1 and p53 as an increase of DNA repair capacity was registered only after silencing of both proteins. PMID:26896489

  5. 淫羊藿苷逆转耐甲氨蝶呤肺癌A549细胞转移表型%Icariin reversed metastatic phenotype of methotrexate-resistant lung cancer A549 cells

    吴剑锋; 何晓东; 许卫东; 李道静; 孙利; 沈佐君

    2009-01-01

    目的:研究中药淫羊藿苷(icariin,ICA)作用甲氨蝶呤(methotrexate,MTX)耐药肺癌A549细胞后对细胞转移表型的影响,初步探讨ICA逆转A549/MTX耐药细胞转移表型的作用机制及对肺癌的治疗价值.方法:采用MTT法检测ICA对A549/MTX耐药细胞的半数抑制浓度(half inhibition concentration,IC_(50)).采用双层软琼脂克隆形成实验检测A549/MTX 组和A549/MTX+ICA组细胞的克隆形成率,并观察其集落形态.细胞划痕实验检测A549/MTX组和A549/MTX+ICA组细胞的迁移能力.Transwell小室实验检测细胞侵袭能力的变化.结果:MTT结果显示,无毒剂量的ICA与MTX联合应用后A549/MTX细胞的IC_(50)值为35.50±1.85 μmol/L,比单独应用MTX(同等剂量)后A549/MTX细胞的IC50值(52.17±2.25 μmol/L)有了一定程度的下降.软琼脂实验发现,A549/MTX+ICA组细胞克隆形成率为0.94±0.09,小于A549/MTX组细胞的1.56±1.07(P<0.05).划痕实验显示,72 h后A549/MTX组细胞的迁移能力大于A549/MTX+ICA组细胞(P<0.05).Transwell实验显示,A549/MTX组细胞的穿膜细胞数明显多于A549/MTX+ICA组细胞(P<0.05),说明A549/MTX+ICA组细胞的侵袭浸润能力小于A549/MTX组细胞.结论:中药ICA具有逆转A549/MTX耐药细胞转移表型的作用.

  6. Mimulone-Induced Autophagy through p53-Mediated AMPK/mTOR Pathway Increases Caspase-Mediated Apoptotic Cell Death in A549 Human Lung Cancer Cells

    An, Hyun-Kyu; Kim, Kyoung-Sook; Lee, Ji-Won; Park, Mi-Hyun; Moon, Hyung-In; Park, Shin-Ji; Baik, Ji-Sue; Kim, Cheorl-Ho; Lee, Young-Choon

    2014-01-01

    Anticancer properties and mechanisms of mimulone (MML), C-geranylflavonoid isolated from the Paulownia tomentosa fruits, were firstly elucidated in this study. MML prevented cell proliferation in a dose- and time-dependent way and triggered apoptosis through the extrinsic pathway in A549 human lung adenocarcinoma cells. Furthermore, MML-treated cells displayed autophagic features, such as the formation of autophagic vacuoles, a primary morphological feature of autophagy, and the accumulation ...

  7. Effect of Gemcitabine in the Uptake of 18F-FDG Non-small-cell on Human Lung Cancer Cell A549%吉西他滨对人非小细胞肺癌A549细胞摄取18F-FDG影响的研究

    邹惠峰; 邓胜明; 章斌; 吴翼伟

    2011-01-01

    目的 探讨测定人非小细胞肺癌A549细胞的18F-FDG细胞结合率方法及用吉西他滨化疗后对A549细胞摄取18FFDG的影响.方法 在不同条件下测定A549细胞的18F-FDG细胞结合率,细胞浓度5×104~1×107/瓶;18F-FDG放射性活度1.85~29.6KBq;反应时间20~120min;葡萄糖浓度0~11.1mmol/L.MTT测定加入不同剂量0~120mmol/L吉西他滨24h后细胞抑制率.测定加入不同剂量0~120mmol/L吉西他滨24h后18F-FDG细胞结合率.结果 18F-FDG细胞结合率随细胞数量、反应时间的增加而增高,随葡萄糖浓度的增高而降低,与18F-FDG放射性活度无关;加入不同剂量吉西他滨后,细胞结合率随剂量增加而下降,两者呈负相关(r=-0.78,P<0.01).结论吉西他滨作用24h后引起人非小细胞肺癌A549细胞 18F-FDG细胞结合率下降,可用18F-FDG显像早期观测吉西他滨对人非小细胞肺癌疗效.%Objective To optimize the measurement of 18F-FDG uptake rates of non-small-cell lung cancer cell A549 and investigate the effect after administrated with Gemcitabine. Methods To detect 18F-FDG uptake rates of non-small-cell lung cancer A549 cells in different conditions:cell density ranges from 5 × 104 to 1 × 107per flask,radioactivity of 18F-FDG from 1.85 to 29.6KBq,incubating time from 20 to 120 minutes,glucose concentration from 0 to 11.1mmol/L.24 hours after administrated with Gemcitabine(0 ~ 120mmol/L),inhibition ratios and 18F-FDG uptake rates of the A549 cells were detected. Results On certain conditions,18F-FDG uptake rates of A549 cells increased as the cell number and incubating time grew,but decreased while glucose concentration raised,irrelative with radioactivity of 18F-FDG.18F-FDG uptake rates of A549 cells decreased with the concentration of Gemcitabine increasing,which presented negative correlation(r=-0.78,P<0.01).Conclusions 18F-FDG uptake rates of non-small-cell lung cancer A549 cells decreased 24 hours after treated with Gemcitabine

  8. A Novel Bufalin Derivative Exhibited Stronger Apoptosis-Inducing Effect than Bufalin in A549 Lung Cancer Cells and Lower Acute Toxicity in Mice

    Liu, Miao; Feng, Li-Xing; Sun, Peng; Liu, Wang; Wu, Wan-Ying; Jiang, Bao-Hong; Yang, Min; Hu, Li-Hong; Guo, De-An; Liu, Xuan

    2016-01-01

    BF211 is a synthetic molecule derived from bufalin (BF). The apoptosis-inducing effect of BF211 was stronger than that of BF while the acute toxicity of BF211 was much lower than that of BF. BF211 exhibited promising concentration-dependent anti-cancer effects in nude mice inoculated with A549 cells in vivo. The growth of A549 tumor xenografts was almost totally blocked by treatment with BF211 at 6 mg/kg. Notably, BF and BF211 exhibited differences in their binding affinity and kinetics to recombinant proteins of the α subunits of Na+/K+-ATPase. Furthermore, there was a difference in the effects of BF or BF211 on inhibiting the activity of porcine cortex Na+/K+-ATPase and in their time-dependent effects on intracellular Ca2+ levels in A549 cells. The time-dependent effects of BF or BF211 on the activation of Src, which was mediated by the Na+/K+-ATPase signalosome, in A549 cells were also different. Both BF and BF211 could induce apoptosis-related cascades, such as activation of caspase-3 and the cleavage of PARP (poly ADP-ribose polymerase) in A549 cells, in a concentration-dependent manner; however, the effects of BF211 on apoptosis-related cascades was stronger than that of BF. The results of the present study supported the importance of binding to the Na+/K+-ATPase α subunits in the mechanism of cardiac steroids and also suggested the possibility of developing new cardiac steroids with a stronger anti-cancer activity and lower toxicity as new anti-cancer agents. PMID:27459387

  9. Biochemical synthesis of silver nanoprticles using filamentous fungi Penicillium decumbens (MTCC-2494) and its efficacy against A-549 lung cancer cell line.

    Majeed, Shahnaz; Abdullah, Mohd Syafiq Bin; Dash, Gouri Kumar; Ansari, Mohammed Tahir; Nanda, Anima

    2016-08-01

    Biosynthesis of silver and other metallic nanoparticles is one of the emerging research area in the field of science and technology due to their potentiality, especially in the field of nano-biotechnology and biomedical sciences in order to develop nanomedicine. In our present study, Penicillium decumbens (MTCC-2494) was brought from Institute of Microbial Technology (IMTECH) Chandigarh and employed for extracellular biological synthesis of silver nanoparticles. Ag-NPs formation was appeared with a dark brown color inside the conical flask. Characterization of Ag-NPs were done by UV-Spectrophotometric analysis which showed absorption peak at 430 nm determines the presence of nanoparticles, Fourier transform infrared (FT-IR) spectroscopic analysis, showed amines and amides are the possible proteins involved in the stabilization of nanoparticles as capping agent. Atomic force Microscopy (AFM) confirmed the particle are spherical, size was around 30 to 60 nm and also the roughness of nanoparticles. Field emission scanning electron microscopy (FE-SEM) showed the topology of the nanoparticles and were spherical in shape. The biosynthesis process was found fast, ecofriendly and cost effective. Nano-silver particle was found to have a broad antimicrobial activity and also it showed good enhancement of antimicrobial activity of Carbenicillin, Piperacillin, Cefixime, Amoxicillin, Ofloxacin and Sparfloxacin in a synergistic mode. These Ag-NPs showed good anti-cancer activity at 80 μg·mL(-1)upon 24 hours of incubation and toxicity increases upon 48 hours of incubation against A-549 human lung cancer cell line and the synergistic formulation of the antibiotic with the synthesized nanoparticles was found more effective against the pathogenic bacteria studied. PMID:27608951

  10. 氨甲蝶呤对映体对肺癌A549细胞的生长抑制作用研究%Inhibitory effects of methotrexate enantiomers on the growth of human lung cancer A549 cells

    陶绍能; 王莹莹; 周建国; 程光华; 张梦莹; 钟民; 吕坤

    2015-01-01

    目的:研究氨甲喋呤(MTX)对映体对肺癌A549细胞的生长抑制作用.方法:倒置相差显微镜观察加入MTX对映体后细胞形态变化,应用MTT法检测MTX对映体对A549细胞的生长抑制作用,应用流式细胞术分析细胞周期分布及凋亡率的变化.结果:倒置相差显微镜观察加入MTX对映体后细胞形态发生明显变化,MTT法检测表明两种MTX对映体抑制A549细胞的生长呈剂量与时间依赖性,L-(+)-MTX对A549细胞的抑制作用明显强于D-(-)-MTX.流式细胞检测发现两种MTX对映体药物对A549细胞的细胞周期和凋亡具有明显干扰作用.结论:MTX对映体对A549细胞的作用明显具有手性差异,L-(+)-MTX对A549细胞的抑制作用明显强于D-(-)-MTX.

  11. Luciferase bioluminescence imaging monitoring gene therapeutic effect of apoptosis-inducing ligand for lung cancer A549 cells nude mice transplantation tumor in vivo

    Objective: To detect the expression and effect of human tumor necrosis factor related apoptosis-inducing ligand (hTRAIL) in vivo,by using a novel double expressing adenoviral vector encoding hTRAIL and firefly luciferase (luc) gene (ad-luc-hTRAIL), in which luc was used as reporter gene. Methods: Lung cancer A549 cell xenografts in 16 nude mice models were established in subcutaneous inoculation way, the adenovirus vectors (ad-luc-hTRAIL, ad-hTRAIL, ad-luc) and phosphate buffer saline (PBS) (n=4) as control were injected into tumor respectively. The size of the tumor was measured at different time points (4, 7, 10, 14, 21, 28 d) after injection. The activity of luciferase in surface of the tumor was detected in vivo by using high-sensitivity cooled-charged coupled device (CCD) camera. The expression of hTRAIL was demonstrated by immunohistochemistry staining after sacrificing the animals at different time points, and immunohistochemical scores (IHS) were measured. The apoptosis rate of tumor cells was detected by using TUNEL and calculated. Analysis of variance, the paired t test and linear correlation analysis was used for the statistics. Results: The growing speed of tumour xenografts was more slowly in ad-luc-hTRAIL and ad-hTRAIL groups than PBS group (t=2.71, 2.72, P<0.05). The tumor volumes of ad-luc-hTRAIL, ad-hTRAIL, ad-luc and PBS groups 28 days after injection were (208.4 ± 42.3), (181.5 ±23.9), (403.1 ± 54.0) and (427.0 ± 59.3) mm3, respectively. There was no significant difference between ad-luc group and PBS group (t=2.07, P>0.05). The expression of luciferase in ad-luc-hTRAIL group reached its peak at 7th day (1.37 ± 1.04), and then decreased quickly. The IHS and apoptosis rate in ad-luc-hTRAIL and ad-hTRAIL groups reached their peaks at 7th day, the peak values of IHS were 6.25 ±2.06 and 6.5 ± 2.89, the peak values of apoptosis rate were (60.75 ± 8.06)% and (61.50 ± 8.47)%,respectively. The amount of luciferase expression (absolute number of

  12. The Study on Anti-cancer Effects of Distilling Fresh-ginseng Herbal acupuncture against implanted Sarcoma-180 in vivo and A549 human epithelial lung cancer cells in vitro

    Hae-Young Jang

    2004-12-01

    Full Text Available Objectives : This study was to investigate the anti-cancer effects of herbal acupuncture with distilled fresh ginseng. The herbal acupuncture was injected to Chung-wan(C.V12 and Wisu(BL21 of mice that were subjected to Sarcoma-180 abdominal cancer cell and A549 human epithelial lung cancer cells in vitro. Methods : Anti-cancer effects of distilled fresh ginseng herbal acupuncture were tested by measruing Cox, Bcl-2, and Bax by using RT-PCR in A549 human epithelial lung cancer cells in vitro. And four weeks old Balb/c line male mice weighing around 20±3g were used to measure survival rate and anti-cancer effect to outputs of interleukin-2 and interleukin-4 using flow cytometry, possibility of mRNA menifestation using RT-PCR, and Cox mRNA. The results are as follows. Results : 1. In measuring mRNA menifestation in Cox, Bcl-2, and Bax by using RT-PCR in A549 human epithelial lung cancer cells in vitro, the result showed that fresh ginseng decreased Cox-2 which is directly involved in inflammation process. 2. Survival rate was measured in an anti-cancer effect experiment against Sarcoma-180 abdorminal cancer. Median survival time of controlled group was 27 days, of experiment group I was 21 days, and of experiment group II was 27 days. Therefore, experiment group I showed -22.2% increase in survival rate and experiment group II showed no difference compare to controlled group. 3. There was no difference between condition group and controlled and experiment group in measuring outputs of interleukin-2 and interleukin 4 by using flow cytometry 4. In measuring outputs of interleukin-2 by using ELISA, there was no significant difference between condition group and controlled group and there was decrease in experiment group II compared to conditioned and controlled group. 5. In measuring cytokine mRNA menifestation by using RT-PCR, experiment group I showed increase of mRNA menifestation in interleukin-2,4 and interferon-γ and experiment group II showed

  13. Phospholipid flippase associates with cisplatin resistance in plasma membrane of lung adenocarcinoma A549 cells

    2001-01-01

    The fusion of the liposomes containing N-(7-nitro-2, 1, 3-benzoxadiazol-4-yl)-i ,2-hexadecanoylSn-glycero-3-1abeled phosphatidylethanolamine (NBD-PE) with A549 and A549/DDP cells was performed, and the activity of the phospholipid flippase in the plasma membrane of the cells was measured by fluorescence intensity change of NBDPE in the outer membrane. When A549 or A549/DDP cells containing N BD-PE were incubated at 37 C for 0, 30, 60 and 90 min, the fluorescence intensities in the outer membrane of the cells were 0%, 1.4%, 2.9% and 7.8% for A59cells, and 0%, 10.5 %, 15. 5 % and 18.3 % for A549/DDP cells respectively, demonstrating that the phospholipid flippase was distributed in the plasma membrane of As49 cells, but its activity in the drug-resistant A549/DDP cells was much higher than that in the A549 cells. When the A549/DDP cells were incubated with a multidrug resistance reverse agent, verapamil, for 60 min at 37C, the results showed that the NBD-PE in outer membrane decreased by 25.0% compared with the control's. Furthermore, when A549/DDP cells were incubated with 25 μmol/L cisplatin, which is a specific anticancer drug, the flippase activity decreased by 31.6%, and it further decreased with the increase of cisplatin concentration, suggesting that phospholipid flippase in the membrane might be related to the cisplatin-resistance of human lung adenocarcinoma cancer cells.

  14. An Experimental Study on Effects of Distilled White-ginseng Herbal Acupuncture on A549 human ephithelial lung cancer cell in vitro and implanted Sarcoma-180 in vivo

    Jong-Seong We; Ki-Rok Kwon; Hee-Soo Park

    2004-01-01

    Objectives : In order to investigate effects and immune improvement of distilled white-ginseng herbal extract, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled white-ginseng Herbal Acupuncture at Wisu(BL21) and Chung-wan(CV12) to investigate anti-cancer effects and immune response. R...

  15. An Experimental Study on Effects of Distilled Red-ginseng Herbal Acupuncture on A549 human ephithelial lung cancer cell in vitro and implanted Sarcoma-180

    Seung Hwan Won; Ki-Rok, Kwon; Sun-Gu, Lee

    2004-01-01

    Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu(BL21) and Chung- wan(CV12) to investigate anti-cancer effects and immune response. ...

  16. Identification and Isolation of Cancer Stem Cells from A549 Cells

    Hui XIA

    2013-08-01

    Full Text Available Background and objective Lung cancer stem cells are the root causes of lung cancer malignant phenotype and potential therapeutic target, the aim of this study is to isolate and characterize the cancer stem cells in the lung adenoearcinomas cell line A549, so as to provide an experimental basis for further stem cell research. Methods The cancer stem cells were isolated from the lung adenoearcinomas cell line A549 using FACS. And the difference of colony formation, cell proliferation and tumorigenicity in vitro were also tested. The expression of CD133 and ABCG2 were evaluated by RT-PCR and Western blot. Results The percentage of SP cells was 5.93% of A549 and 0.32% of A549 after incubation with verapamil. The results showed that there were significantly higher expression of CD133 and ABCG2 on SP cells than that of non-SP cells. And the ability of colony formation, cell proliferation and tumorigenicity in SP cell group were remarkably higher than that in non-SP cell group. Conclusion Our results suggested that the cancer stem cells with higher expression of CD133 and ABCG2 can be isolated from the lung adenoearcinomas cell line A549 using FACS and be used in the further research experiments.

  17. A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

    Highlights: ► A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. ► We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. ► The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. ► The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

  18. A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

    Lu, Xiangyi; Liu, Wei; Wu, Junhua; Li, Mengxian [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China); Wang, Juncheng; Wu, Jihui [School of Life Science, University of Science and Technology of China, Hefei 230022 (China); Luo, Cheng, E-mail: Luo58@yahoo.com [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China)

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. Black-Right-Pointing-Pointer We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. Black-Right-Pointing-Pointer The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. Black-Right-Pointing-Pointer The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

  19. Sulforaphane derived from broccoli inhibit proliferation and invasion of lung cancer A549 cells in vitro%西兰花提取物萝卜硫素抑制肺癌细胞的生长和侵袭

    贾侃; 贺云冲; 洪姣; 黄春琦; 任军; 许健

    2014-01-01

    Sulforaphane was a multifunction compound derived from brassicaceous vegetable such as broccoli, reports showed that Sulforaphane provided with effection of antitumor and antioxidant. Lung cancer is an aggressive malignancy with a tendency of early distant metastases, the antitumor function of sulforaphane was corroborated by numerous lines of evidence, but the anticancer mechanism of this compound has not been wel obsvered. In this work, we analyzed vitality and invasion of A549 cels treated with sulforaphane by cellcounting kit (CCK8) and transwel, then measure the half maximal (50%) inhibitory concentration (IC50) of sulforaphane for A549 cels. The cels cycle, apoptosis and DNA fragment were analyzed using Flow Cytometry Analysis and agarose electrophoresis, TGF-βand NF-κB were analyzed by western blot after treatment with 3μg/mL sulforaphane. Results showed that A549 cels proliferate and invade were inhibited by sulforaphane with a dose-dependent manner, IC50 of sulforaphane was 3μg/mL, and the cellcycle were arrested at G2/M phase. 3μg/mL sulforaphane induced apoptosis , DNA fragment, decreased the expression of TGF-βand NF-κB in A549 cels. Our results pointed out that sulforaphane inhibited proliferation and invasion of lung cancer A549 cels in vitro, decreased the expression of inflammation proteins, maybe a novel chemotherapy for lung cancer.%萝卜硫素是从十字花科蔬菜中提取的多功能物质,研究已证实其具有抗癌、抗氧化等功效。肺癌是恶性程度高、具有转移倾向的恶性肿瘤,萝卜硫素抗肺癌的机制尚不是十分清楚。本研究通过CCK-8和transwel侵袭实验分析初步判断萝卜硫素对A549肺癌细胞活性和转移侵袭的影响,计算体外干预A549的IC50,流式细胞学分析IC50浓度萝卜硫素对细胞周期和凋亡的影响,电泳分析DNA片段化改变。结果显示A549细胞活性对萝卜硫素剂量依赖性下降,萝卜硫素作用于A549细胞的IC50为3μg

  20. Effects of 5-Aza-Cde on DNA Methylation and Expression of hMLHl and MGMT Gene in Lung Cancer Cell Line A549/DDP%5-氮杂-2′脱氧胞苷对肺癌 A549/DDP 细胞hMLHl,MGMT 基因甲基化及其表达的影响

    王虹; 李丽丽; 张吉才; 高波; 骆海军

    2015-01-01

    Objective To investigate the effects of 5-Aza-2′-deoxycytidine (5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific pol-ymerase chain reaetioll (MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression of hMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively. Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter region of the hM-LH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.%目的:观察5-氮杂-2′脱氧胞苷(5-Aza-Cde)对体外培养的顺铂(DDP)耐药株肺癌 A549/DDP 细胞 hMLH1,MG-MT 基因启动子区 DNA 甲基化状态及其表达的影响,探讨肺癌细胞 hMLH1和 MGMT 基因失活的机制及去甲基化制剂对 hMLH1和 MGMT 基因表达的调控。方法5-Aza-Cde 处理体外1640培养的肺癌 A549/DDP 细胞,甲基化特异性PCR(MSP)法检测用药前后细胞 hMLH1和 MGMT 基因的甲基化状态,RT-PCR 法检测用药前后细胞 hMLH1和 MG-MT mRNA 的表达。结果在对照组 A549细胞当中 hMLH1基因是非甲基化状态和高表达,而 MGMT 显示为低甲基化(部分甲基化)状态和高表达;而在顺铂耐药株 A549-DDP 中,hMLH1和 MGMT 基因均显示高甲基化状态,mRNA 表达下调。结论hMLH1和 MGMT 基因甲基化修饰程度与 mRNA 的表

  1. Influence of suppressor gene p16 on retinoic acid inducing cancer cell A549 differentiation

    2001-01-01

    Objective To investigate the role of suppressor gene p16 in the process of differential regulation of retinoic acid (RA) on the A549 lung cancer cells.Methods Tumor suppressor gene p16 was transferred into A549 cells and the cells were treated with all-trans retinoic acid (ATR) at the dosage of 5×10-6 mol/L for 4 d. After that, the proliferation and differentiation of A549 cells were examined by growth curve and cytometry analysis, the change of lung lineage-specific marker MUC1 was tested by immunohistochemical staining. Meanwhile, Western blot was used to observe the change of p16 protein expression in A549 cells treated with ATRA.Results ATRA could obviously inhibit the growth and induce the differentiation of A549 Cells that were transferred with p16 gene. There were more cells arrested in G1/G0 phase and the expression of MUG1 was markedly down-regulated than in control cells. The expression of p16 protein was up-regulated in A549 cells treated with ATRA.Conclusion Suppressor gene p16 could enhance the effects of RA and proliferated suppression and differential induction of A549 cells.

  2. All-Trans Retinoic Acid Induces Proliferation, Survival, and Migration in A549 Lung Cancer Cells by Activating the ERK Signaling Pathway through a Transcription-Independent Mechanism

    Quintero Barceinas, Reyna Sara; García-Regalado, Alejandro; Aréchaga-Ocampo, Elena; Villegas-Sepúlveda, Nicolás; González-De la Rosa, Claudia Haydée

    2015-01-01

    All-trans retinoic acid (ATRA) has been used as an antineoplastic because of its ability to promote proliferation, inhibition, and differentiation, primarily in leukemia; however, in other types of cancer, such as lung cancer, treatment with ATRA is restricted because not all the patients experience the same results. The ERK signaling pathway is dysregulated in cancer cells, including lung cancer, and this dysregulation promotes proliferation and cell invasion. In this study, we demonstrate that treatment with ATRA can activate the ERK signaling pathway by a transcription-independent mechanism through a signaling cascade that involves RARα and PI3K, promoting growth, survival, and migration in lung cancer cells. Until now, this mechanism was unknown in lung cancer cells. The inhibition of the ERK signaling pathway restores the beneficial effects of ATRA, reduces proliferation, increases apoptosis, and blocks the cell migration process in lung cancer cells. In conclusion, our results suggest that the combination of ATRA with ERK inhibitor in clinical trials for lung cancer is warranted. PMID:26557664

  3. SchA-p85-FAK complex dictates isoform-specific activation of Akt2 and subsequent PCBP1-mediated post-transcriptional regulation of TGFβ-mediated epithelial to mesenchymal transition in human lung cancer cell line A549.

    Xue, Xinying; Wang, Xin; Liu, Yuxia; Teng, Guigen; Wang, Yong; Zang, Xuefeng; Wang, Kaifei; Zhang, Jinghui; Xu, Yali; Wang, Jianxin; Pan, Lei

    2014-08-01

    A post-transcriptional pathway by which TGF-β modulates expression of specific proteins, Disabled-2 (Dab2) and Interleukin-like EMT Inducer (ILEI), inherent to epithelial to mesenchymal transition (EMT) in murine epithelial cells through Akt2-mediated phosphorylation of poly r(C) binding protein (PCBP1), has been previously elucidated. The aims of the current study were to determine if the same mechanism is operative in the non-small cell lung cancer (NSCLC) cell line, A549, and to delineate the underlying mechanism. Steady-state transcript and protein expression levels of Dab2 and ILEI were examined in A549 cells treated with TGF-β for up to 48 h. Induction of translational de-repression in this model was quantified by polysomal fractionation followed by qRT-PCR. The underlying mechanism of isoform-specific activation of Akt2 was elucidated through a combination of co-immunoprecipitation studies. TGF-β induced EMT in A549 cells concomitant with translational upregulation of Dab2 and ILEI proteins through isoform-specific activation of Akt2 followed by phosphorylation of PCBP1 at serine-43. Our experiments further elucidated that the adaptor protein SchA is phosphorylated at tyrosine residues following TGF-β treatment, which initiated a signaling cascade resulting in the sequential recruitment of p85 subunit of PI3K and focal adhesion kinase (FAK). The SchA-FAK-p85 complex subsequently selectively recruited and activated Akt2, not Akt1. Inhibition of the p85 subunit through phosphorylated 1257 peptide completely attenuated EMT in these cells. We have defined the underlying mechanism responsible for isoform-specific recruitment and activation of Akt2, not Akt1, during TGF-β-mediated EMT in A549 cells. Inhibition of the formation of this complex thus represents an important and novel therapeutic target in metastatic lung carcinoma. PMID:24819169

  4. Determination of in vitro free radical scavenging and antiproliferative effect of Pennisetum alopecuroides on cultured A549 human lung cancer cells

    Githa Elizabeth Mathew

    2015-01-01

    Conclusions: This is the 1 st time a pharmacological exploration of P. alopecuroides grasses has been conducted. We have shown that P. alopecuroides exhibits good free radical scavenging and strong in vitro cytotoxic activities against human lung cancer cell lines.

  5. Radix Tetrastigma hemsleyani flavone inhibits proliferation, migration, and invasion of human lung carcinoma A549 cells

    Zhong LR

    2016-02-01

    Full Text Available Liangrui Zhong,1 Junxian Zheng,2 Qianqian Sun,3 Kemin Wei,2 Yijuan Hu2 1Department of Oncology, Tongde Hospital of Zhejiang Province, Affiliated to Zhejiang Chinese Medical University, 2Department of Chinese Medicine, Zhejiang Academy of Traditional Chinese Medicine, 3Department of Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China Abstract: Radix Tetrastigma hemsleyani flavone (RTHF is widely used as a traditional herb and has detoxification and anti-inflammatory effects. In this study, we investigated the potential effects of RTHF on the growth and metastasis of human lung adenocarcinoma A549 cells and evaluated its mechanisms. A549 cells were treated with RTHF at various concentrations for different periods. In vitro Cell Counting Kit-8 assay and colony formation methods showed that RTHF had dose- and time-dependent antiproliferation effects on A549 cells. A cell adhesion assay showed that RTHF decreased A549 cell adhesion in a dose-dependent manner. Cell invasion and migration were investigated using the Transwell assay and observed using an inverted microscope; the results showed that cell metastasis was significantly lower in the treatment group than that in the control group (P<0.01. Expression of metastasis-related matrix metalloproteinases (MMPs and tissue inhibitors of metalloproteinases (TIMPs was detected by real-time polymerase chain reaction and Western blotting. The results showed that the expression of MMP-2, MMP-9, and TIMP-1 decreased, while that of TIMP-2 increased significantly in the RTHF group when compared with the results of the control group. These results show that RTHF exhibits antigrowth and antimetastasis activity in lung cancer A549 cells by decreasing the expression of MMP-2/-9 and TIMP-1 and increasing that of TIMP-2. Keywords: flavone, radix Tetrastigma hemsleyani, metastasis, lung cancer

  6. Effect of antisense transfecting of monocarboxylate transporter gene on biological characteristics of lung adenocarcinoma A549 cells

    ZHANG Gui-zhi; HUANG Gui-jun; GUO Xian-jian; QIAN Gui-sheng

    2002-01-01

    Objective: To study the influence of transfecting antisense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into lung cancer cells on pHi regulation, lactate transportation and cell growth, Methods: MCT1 antisense gene recombinant vector was introduced into human lung cancer cell line A549 by electroporation. The transfected A549 cells resistant to G418 were selected. Positive clones were examined by using PCR. The changes of intracellular pH and lactate were examined with spectrophotometric method. Cell growth was studied with cell growth curve. Results: Intracellular pH and lactate were remarkably decreased in the cells transfected pLXSN-MCT1 in comparison with A549 cells without transfection (P<0. 001). The growth of A549 cells transfected pLXSN-MCT1 was also inhibited remarkably. Conclusion: MCT1 gene may play an important role in pHi regulation, lactate transportation and cell growth in tumor cells.

  7. Effect of fucoidan from Turbinaria conoides on human lung adenocarcinoma epithelial (A549) cells.

    Alwarsamy, Madhavarani; Gooneratne, Ravi; Ravichandran, Ramanibai

    2016-11-01

    Fucoidan was purified from seaweed, Turbinaria conoides. Isolated fragments were characterized with NMR ((13)C, (1)H), Gas Chromatography-Mass Spectronomy (GC-MS) and HPLC analysis. The autohydrolysate of fucoidans consisted of sulfated fuco-oligosaccharides having the backbone of α-(1, 3)-linked fuco-pyranose derivatives and minor components of galactose, glucose, mannose and xylose sugars. Fucoidan induced a dose-dependent reduction in cell survival of lung cancer A549 cells by MTT assay (GI50, 75μg/mL). However, it was not cytotoxic to a non-tumorigenic human keratinocyte cell line of skin tissue (HaCaT) (GI50>1.0mg/mL). The apoptotic cells in fucoidan-treated A549 cells were visualized by laser confocal microscopy and cell cycle analysis showed induction of G0/G1 phase arrest of the cell progression cycle. Further, CFSE labeling and flow cytometry highlighted that fucoidan significantly (P<0.05) inhibited the proliferation rate of A549 cells by up to 2-fold compared with the control cells. It is concluded that fucoidan has the potential to act as an anti-proliferative agent on lung carcinoma (A549) cells. PMID:27516266

  8. Sodium fluoride toxicity and its combined effect with 12C-ion beam radiation on A549 lung cancer cell line

    Sodium fluoride (NaF) is an anti-glycolytic agent (inhibits enolase) thus blocking formation of pyruvate. Not much has been documented about its radio-sensitization properties. We are trying to explore its synergism with high LET radiation to kill the cancer cells. We have determined the IC50 of NaF in A549 cell line to be 13.7 mM. A minimum dose of 1mM NaF was selected on the basis of proliferation, morphological, cell cycle studies, to be used in combination with a 12C-ion beam dose of 1 Gy. NaF is administered for 24 h prior to irradiation and then assayed for sensitivity after 24 h of post irradiation incubation. Cell growth decreased in the combination treatment in comparison to radiation alone. Cell cycle analysis reveals G2-block upon 1 Gy irradiation in addition to the DNA fragmentation in the combination treatment. Metabolic activity as measured by MTT assay increased gradually along with ROS as evident in DCFDA assay. An important observation is the significant reduction in mitochondrial membrane potential in the combination treatment compared to radiation treatment alone as observed in total fluorescence of Rhodamine 123. We, hereby, aim to minimize the radiation dose by synergizing with optimum dose of NaF which would contribute to more efficient and economical heavy ion therapy protocols. We have been able to note sensitization in A549. The mitochondrial sensitivity obtained is significant and therefore other cellular event that may lead to cell death at the given dose, therefore signifying the use of high-LET radiation, needs further exploration and is therefore under investigation. (author)

  9. Ent-11α-Hydroxy-15-oxo-kaur-16-en-19-oic-acid Inhibits Growth of Human Lung Cancer A549 Cells by Arresting Cell Cycle and Triggering Apoptosis

    Li Li; George G Chen; Ying-nian Lu; Yi Liu; Ke-feng Wu; Xian-ling Gong; Zhan-ping Gou; Ming-yue Li; Nian-ci Liang

    2012-01-01

    Objective:To examine the apoptotic effect of ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F),a compound isolated from Pteris semipinnata L(PsL),in human lung cancer A549 cells.Methods:A549 cells were treated with 5F (0-80 μg/ml) for different time periods.Cytotoxicity was examined using a MTT method.Cell cycle was examined using propidium iodide staining.Apoptosis was examined using Hoechst 33258 staining,enzyme-linked immunosorbent assay (ELISA) and caspase-3 activity analysis.Expression of representative apoptosis-related proteins was evaluated by Western blot analysis.Reactive oxygen species (ROS) level was measured using standard protocols.Potential interaction of 5F with cisplatin was also examined.Results:5F inhibited the proliferation of A549 cells in a concentration- and time-dependent manner.5F increased the accumulation of cells in sub-G1 phase and arrested the cells in the G2 phase.Exposure to 5F induced morphological changes and DNA fragmentation that are characteristic of apoptosis.The expression of p21 was increased.5F exposure also increased Bax expression,release of cytochrome c and apoptosis inducing factor (AIF),and activation of caspase-3.5F significantly sensitized the cells to cisplatin toxicity Interestingly,treatment with 5F did not increase ROS,but reduced ROS production induced by cisplatin.Conclusion:SF could inhibit the proliferation of A549 cells by arresting the cells in G2 phase and by inducing mitochondrial-mediated apoptosis.

  10. 依维莫司对人非小细胞肺癌细胞系A549放射增敏作用%Effect of Everolimus on Radiosensitivity of Human Non_small Cell Lung Cancer Cell Line A549

    陈豫; 褚倩; 郭娟; 黄玉; 李文雯; 田逸俊; 夏曙; 于世英

    2014-01-01

    目的:通过使用哺乳动物雷帕霉素靶蛋白mTOR抑制药依维莫司抑制A549细胞mTOR信号通路,研究依维莫司是否具有放射增敏作用。方法单纯放射治疗(放疗)或联合依维莫司作用于人非小细胞肺癌细胞系A549,采用噻唑蓝( MTT)法测定依维莫司对A549细胞抑制率并计算半数抑制浓度( IC50)。应用药物20%抑制浓度( IC20)作用24 h后X线2,4,6,8 Gy照射。计算细胞克隆存活分数及多靶单击模型拟合生存曲线,并计算平均致死剂量( D0)、准阈剂量(Dq)、照射剂量2 Gy下细胞存活分数(SF2)和放射增敏比(SER)。采用Western blot 方法检测γ_H2AX蛋白的表达,并分析相对灰度值。结果依维莫司联合放疗可明显提高A549细胞对射线的敏感性,依维莫司+照射组D0、Dq及SF2均明显低于单纯照射组,SER为1.36。依维莫司+照射组X线照射后24 h点γ_H2AX蛋白残余量明显高于单纯照射组。结论依维莫司抑制mTOR信号通路能够提高A549细胞的放射敏感性。%Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was

  11. The mechanism of CpG ODN enhancing the radiosensitivity of lung cancer cell line A549%CpG ODN增强人肺癌细胞株A549放射增敏作用的研究

    颜伟; 孙梯业; 杨春敏; 贾敏; 李静; 史蕊; 唐和兰; 杜斌; 韩全利

    2011-01-01

    Objective To investigate the effect of CpG ODN on the radiosensitivity of lung epithelial cell line A549.Methods The TNF-α,IL-12 and INF-γ secretion by A549 were detected by ELISA;NO level was tested by Griess method ,AP-1 activation within A549 cells was observed using electrophoretic mobility shift assay.Results The inhibitory role was enhanced when CpG ODN 1826(10μg/ml)were combined with β-ray irradiation ,with the increase of TNF-α,IL-2 and INF - γ secretion by cells.CpG ODN1826 combined with β-ray irradiation increased NO leve in A549 cells and inhibited the AP-1 activation within A549 cells.Conclusions CpG ODN1826 can increase the radiosensitivity of lung epithelial cell line A549 and may be tightly related to increasing secretions of IL-12,IFN-γ,TNF-α and NO from cells and the inhibition of AP-1 activation.%目的 初步探讨CpG ODN增强人肺腺上皮细胞株A549放射增敏作用.方法 ELISA法检测细胞TNF-α、IL-12和INF-γ的分泌水平,Griess检测细胞NO的含量并观察CpGODN1826与β射线诱导A549细胞AP-1活化的抑制作用.结果 CpG ODN增加了人肺癌细胞株A549 TNF-α、IL-12、INF-γ和NO的分泌,在联合β射线照射后对A549细胞的杀伤作用更加显著,并显著抑制A549细胞AP-1的活化.结论 CpG ODN对A549有明显的放射增敏作用,其机制可能与CpG ODN增强+4549细胞分泌TNF-α、IL-L2、INF-γ、NO和抑制A549细胞AP-1的活化有关.

  12. Hyaluronic acid-fabricated nanogold delivery of the inhibitor of apoptosis protein-2 siRNAs inhibits benzo[a]pyrene-induced oncogenic properties of lung cancer A549 cells

    Lin, Chung-Ming; Kao, Wei-Chien; Yeh, Chun-An; Chen, Hui-Jye; Lin, Shinn-Zong; Hsieh, Hsien-Hsu; Sun, Wei-Shen; Chang, Chih-Hsuan; Hung, Huey-Shan

    2015-03-01

    Benzo[a]pyrene (BaP), a component of cooking oil fumes (COF), promotes lung cancer cell proliferation and survival via the induction of inhibitor of apoptosis protein-2 (IAP-2) proteins. Thus knockdown of IAP-2 would be a promising way to battle against lung cancer caused by COF. Functionalized gold nanoparticle (AuNP) is an effective delivery system for bio-active materials. Here, biocompatible hyaluronic acid (HA) was fabricated into nanoparticles to increase the target specificity by binding to CD44-over-expressed cancer cells. IAP-2-specific small-interfering RNA (siRNAs) or fluorescein isothiocyanate (FITC) were then incorporated into AuNP-HA. Conjugation of IAP-2 siRNA into AuNPs-HA was verified by the UV-vis spectrometer and Fourier transform infrared spectrometer. Further studies showed that AuNP-HA/FITC were effectively taken up by A549 cells through CD44-mediated endocytosis. Incubation of BaP-challenged cells with AuNP-HA-IAP-2 siRNAs silenced the expression of IAP-2, decreased cell proliferation and triggered pronounced cell apoptosis by the decrease in Bcl-2 protein and the increase in Bax protein as well as the active form of caspases-3. The BaP-elicited cell migration and enzymatic activity of the secreted matrix metalloproteinase-2 were also substantially suppressed by treatment with AuNP-HA-IAP-2 siRNAs. These results indicated that IAP-2 siRNAs can be efficiently delivered into A549 cells by functionalized AuNP-HA to repress the IAP-2 expression and BaP-induced oncogenic events, suggesting the potential therapeutic application of IAP-2 siRNA or other siRNA-conjugated AuNP-HA composites to COF-induced lung cancer and other gene-caused diseases in the future.

  13. Melatonin inhibits the migration of human lung adenocarcinoma A549 cell lines involving JNK/MAPK pathway.

    Qiaoyun Zhou

    Full Text Available OBJECTIVE: Melatonin, an indolamine produced and secreted predominately by the pineal gland, exhibits a variety of physiological functions, possesses antioxidant and antitumor properties. But, the mechanisms for the anti-cancer effects are unknown. The present study explored the effects of melatonin on the migration of human lung adenocarcinoma A549 cells and its mechanism. METHODS: MTT assay was employed to measure the viability of A549 cells treated with different concentrations of melatonin. The effect of melatonin on the migration of A549 cells was analyzed by wound healing assay. Occludin location was observed by immunofluorescence. The expression of occludin, osteopontin (OPN, myosin light chain kinase (MLCK and phosphorylation of myosin light chain (MLC, JNK were detected by western blots. RESULTS: After A549 cells were treated with melatonin, the viability and migration of the cells were inhibited significantly. The relative migration rate of A549 cells treated with melatonin was only about 20% at 24 h. The expression level of OPN, MLCK and phosphorylation of MLC of A549 cells were reduced, while the expression of occludin was conversely elevated, and occludin located on the cell surface was obviously increased. The phosphorylation status of JNK in A549 cells was also reduced when cells were treated by melatonin. CONCLUSIONS: Melatonin significantly inhibits the migration of A549 cells, and this may be associated with the down-regulation of the expression of OPN, MLCK, phosphorylation of MLC, and up-regulation of the expression of occludin involving JNK/MAPK pathway.

  14. An Experimental Study on Effects of Distilled White-ginseng Herbal Acupuncture on A549 human ephithelial lung cancer cell in vitro and implanted Sarcoma-180 in vivo

    Jong-Seong We

    2004-12-01

    Full Text Available Objectives : In order to investigate effects and immune improvement of distilled white-ginseng herbal extract, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled white-ginseng Herbal Acupuncture at Wisu(BL21 and Chung-wan(CV12 to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups show significant increase. For Cox-2, both experiment groups and the normal group showed significant decrease. For Bcl-2, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 2.For survival time, all of experiment groups didn't show significant differences. 3.For IL-2 productivity using Flow cytometry, experiment group I didn't show any significance, For IL-4, all of experiment groups showed slight decrease compared to the control group. 4. For IL-2 productivity using ELISA, experiment groupI showed slight decrease compared to the control group, experiment group II didn't show any significance. 5.For expression of cytokine mRNA using RT-PCR, significant increase of IL-2 and IL-4 were witnessed in the experiment groupI compared to the control group. Significant decrease of IL-10 was shown in all of experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled white-ginseng Herbal Acupuncture may be further effects in anti-cancer and immune improvement if increasing concentration.

  15. An Experimental Study on Effects of Distilled Red-ginseng Herbal Acupuncture on A549 human ephithelial lung cancer cell in vitro and implanted Sarcoma-180

    Seung Hwan Won

    2004-06-01

    Full Text Available Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu(BL21 and Chung- wan(CV12 to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant decrease. 2.For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3.For survival time, all of experiment groups showed 11.1 % increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5.For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6.For expression of cytokine mRNA using RT-PCR, significant increase of IL-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all of experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effects in anti-cancer and immune improvement if increasing concentration.

  16. Biological evaluation of new nickel(II) metallates: Synthesis, DNA/protein binding and mitochondrial mediated apoptosis in human lung cancer cells (A549) via ROS hypergeneration and depletion of cellular antioxidant pool.

    Kalaivani, P; Saranya, S; Poornima, P; Prabhakaran, R; Dallemer, F; Vijaya Padma, V; Natarajan, K

    2014-07-23

    A series of novel nickel(II) thiosemicarbazone complexes(1-4) have been prepared and characterized by various spectral, analytical techniques and X-ray crystallography. Further, their efficacy to interact with CT-DNA/BSA has been explored. From the binding studies, it is inferred that complex 4 found to be more active than other complexes. The complexes bound with CT-DNA by intercalation mode. Moreover, static quenching was observed for their interaction with BSA. The new complexes were tested for their in vitro cytotoxicity against human lung adenocarcinoma (A549) cell line. The results showed that the new complexes exhibited significant degree of cytotoxicity at given experimental condition. Further, the results of LDH and NO release supported the cytotoxic nature of the complexes. The observed cytotoxicity of the complexes may be routed through ROS-hypergeneration and lipid-peroxidation with subsequent depletion of cellular antioxidant pool (GSH, SOD, CAT, GPx and GST) resulted in the reduction of mitochondrial-membrane potential, caspase-3 activation and DNA fragmentation. Thus, the data from the present study disclose that the complexes could induce apoptosis in A549 cells through mitochondrial mediated fashion and inhibited the migration of lung cancer cells and by metastasis. PMID:24946146

  17. Effects of EPO Gene on Growth and Apoptosis of Lung Adenocarcinoma Cell Line A549

    Jianqing WU

    2009-09-01

    Full Text Available Background and objective Published data on the association between erythropoietin (EPO and cancer cell are inconclusive. The aim of this study is to investigate the effect of erythropoietin (EPO on the growth and survival of lung adenocarcinoma cell line A549. Methods The recombinant plasmid pcDNA3.1(--hEPO was constructed and transfected into A549 cells by liposome protoco1. The Levels of EPO in culture supernatant were detected by ELISA. Effects of EPO gene on growth and survival of the transfected cells were evaluated by MTT assay and flow cytometry (FCM . Levels of vascular endothelial growth factor (VEGF were also evaluated by ELISA. Results The recombinant eukaryotic expression vector pcDNA3.1(--hEPO was successfully constructed. The growth of cells in hEPO transfected cells was significantly inhibited after transfection (P < 0.01. More cells were blocked in S phase in hEPO transfected group compared with control group (P < 0.05, and the apoptotic rate were also significantly higher than those of their controls (P < 0.01. Levels of VEGF in hEPO transfected cells were significantly lower than controls (P < 0.01. Conclusion Exogenous EPO gene expression in A549 cells can induce cell growth inhibition and apoptosis of A549 cells, and expression of VEGF can also be inhibited.

  18. Proteomic response to 5,6-dimethylxanthenone 4-acetic acid (DMXAA, vadimezan in human non-small cell lung cancer A549 cells determined by the stable-isotope labeling by amino acids in cell culture (SILAC approach

    Pan ST

    2015-02-01

    Full Text Available Shu-Ting Pan,1,* Zhi-Wei Zhou,2,3,* Zhi-Xu He,3 Xueji Zhang,4 Tianxin Yang,5 Yin-Xue Yang,6 Dong Wang,7 Jia-Xuan Qiu,1 Shu-Feng Zhou2 1Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People’s Republic of China; 2Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA; 3Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center and Sino-US Joint Laboratory for Medical Sciences, Guiyang Medical University, Guiyang, 4Research Center for Bioengineering and Sensing Technology, University of Science and Technology Beijing, Beijing, People’s Republic of China; 5Department of Internal Medicine, University of Utah and Salt Lake Veterans Affairs Medical Center, Salt Lake City, UT, USA; 6Department of Colorectal Surgery, General Hospital of Ningxia Medical University, Yinchuan, 7Cancer Center, Daping Hospital and Research Institute of Surgery, Third Military Medical University, Chongqing, People’s Republic of China *These two authors contributed equally to this work Abstract: 5,6-Dimethylxanthenone 4-acetic acid (DMXAA, also known as ASA404 and vadimezan, is a potent tumor blood vessel-disrupting agent and cytokine inducer used alone or in combination with other cytotoxic agents for the treatment of non-small cell lung cancer (NSCLC and other cancers. However, the latest Phase III clinical trial has shown frustrating outcomes in the treatment of NSCLC, since the therapeutic targets and underlying mechanism for the anticancer effect of DMXAA are not yet fully understood. This study aimed to examine the proteomic response to DMXAA and unveil the global molecular targets and possible mechanisms for the anticancer effect of DMXAA in NSCLC A549 cells using a stable-isotope labeling by amino acids in cell culture (SILAC approach. The proteomic data showed that treatment with DMXAA

  19. TGF-β1 downregulates COX-2 expression leading to decrease of PGE2 production in human lung cancer A549 cells, which is involved in fibrotic response to TGF-β1.

    Erina Takai

    Full Text Available Transforming growth factor-ß1 (TGF-β1 is a multifunctional cytokine that is involved in various pathophysiological processes, including cancer progression and fibrotic disorders. Here, we show that treatment with TGF-β1 (5 ng/mL induced downregulation of cyclooxygenase-2 (COX-2, leading to reduced synthesis of prostaglandin E2 (PGE2, in human lung cancer A549 cells. Treatment of cells with specific inhibitors of COX-2 or PGE2 receptor resulted in growth inhibition, indicating that the COX-2/PGE2 pathway contributes to proliferation in an autocrine manner. TGF-β1 treatment induced growth inhibition, which was attenuated by exogenous PGE2. TGF-β1 is also a potent inducer of epithelial mesenchymal transition (EMT, a phenotype change in which epithelial cells differentiate into fibroblastoid cells. Supplementation with PGE2 or PGE2 receptor EP4 agonist PGE1-alcohol, as compared with EP1/3 agonist sulprostone, inhibited TGF-β1-induced expression of fibronectin and collagen I (extracellular matrix components. Exogenous PGE2 or PGE2 receptor agonists also suppressed actin remodeling induced by TGF-β1. These results suggest that PGE2 has an anti-fibrotic effect. We conclude that TGF-β1-induced downregulation of COX-2/PGE2 signaling is involved in facilitation of fibrotic EMT response in A549 cells.

  20. 非小细胞肺癌A549细胞摄取18F-FDG早期评价放疗疗效的实验研究%The Experimental Study on Evaluating the Effect of Radiotherapy in the Early Stage by Uptake of 18F-FDG in Human Non-small-cell Lung Cancer A549 Cells

    夏青; 张玮; 章斌; 邓胜明; 吴翼伟

    2012-01-01

    目的 利用18F-脱氧葡萄糖(FDG)细胞结合率的变化来早期评价非小细胞肺癌的放疗疗效.方法 在不同条件下测定非小细胞肺癌A549细胞的18F-FD G细胞结合率,细胞数量为0.5×105~5×106/孔,反应时间为20~120min.对非小细胞肺癌A549细胞进行单纯照射,测定照射后24h和48h18F-FDG细胞结合率,采用四甲基偶氮唑盐(MTT)法测定不同放射剂量作用于A549细胞24h和48h后OD值并计算细胞生长抑制率.结果 当每孔为1 ×106个细胞、加入3.7KBq 18F-FDG、作用时间为100min时,细胞结合率可达(42.96±1.21)%.照射后24h,各剂量组间18F-FDG细胞结合率差异无统计学意义(P> 0.05);照射后48h,各剂量组间18F-FDG细胞结合率随照射剂量的增加而降低,差异有统计学意义(P<0.05);48h后,MTT细胞生长抑制率与18F-FDG细胞结合抑制率呈正相关(r=0.832,P<0.01).结论 单纯照射后48h可引起非小细胞肺癌A549细胞18F-FDG细胞结合率下降,18F-FDG显像有望作为早期评价放疗疗效对非小细胞肺癌敏感性的评价标准之一.%Objective To evaluate the effect of radiotherapy early by uptake of 18F-FDG in human non-small-cell lung cancer A549 cells.Methods The binding efficiency of 18F-FDG was measured under diverse conditions:0.5 × 105~5×106 cells,3.7KBq 18F-FDG,20~120min incubation in 37℃.The human non-small-cell lung cancer A549 cells were exposed to a single fraction of X-ray radiation.The uptake rates of 18F-FDG were calculated at 24 hours and 48 hours after irradiation.Results The binding efficiency was (42.96 ± 1.21)% at the optimum binding condition 1 × 106cells,3.7KBq 18F-FDG and l00min incubation in 37℃.At 24 hours after irradiation,the differences of 18F-FDG uptake rates between groups of various dose were no significant(P>0.05). At 48 hours after irradiation,the 18F-FDG uptake rates decreased with the increasing dose of X-ray in different groups(P<0.05).At 48 hours after irradiation,the binding

  1. Nimesulide acts synergistically with ionizing radiation against A549 human lung cancer cells through the activation of caspase-8 and caspase-3

    Hong, Sung Hee; Kim, Byeong Mo; Maeng, Kyung Ah [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2009-05-15

    Radiotherapy is important in the treatment of non-small cell lung cancer, but very few malignancies have been cured using single modalities of radiotherapy. Therefore, molecules that can target specific pathophysiological or molecular pathways have been investigated for use as radiation sensitizers. Cyclooxygenase (COX)-2 inhibitors have been shown to enhance the radioresponse of cultured human cancer cell lines and immunodeficient mice. However, little is known about the molecular and biochemical mechanisms by which COX-2-selective non-steroidal anti-inflammatory drugs (NSAIDs) enhance the radioresponse of tumor cells. In some types of cancer, radiation is thought to work by inducing apoptosis, and effective anticancer radiotherapy is frequently associated with increased levels of apoptosis markers in vitro and in vivo.

  2. Radiosensitizing Effect of Schinifoline from Zanthoxylum schinifolium Sieb et Zucc on Human Non-Small Cell Lung Cancer A549 Cells: A Preliminary in Vitro Investigation

    Cheng-Fang Wang

    2014-12-01

    Full Text Available Schinifoline (SF, a 4-quinolinone derivative, was found in Zanthoxylum schinifolium for the first time. 4-Quinolinone moieties are thought to have cytotoxic activity and are often used as a tubulin polymerization inhibitors, heterogeneous enzyme inhibitors and antiplatelet agents. However, very little information respect to radiosensitization has focused on SF. This work aimed to investigate the radiosensitizing effect of SF on A549 cells. The cell viability results indicated cytotoxicity of SF on A549 cells, with IC50 values of 33.7 ± 2.4, 21.9 ± 1.9 and 16.8 ± 2.2 μg/mL, respectively, after 6, 12, 24 h treatment with different concentrations, and the 10% or 20% IC50 concentration during 12 h was applied in later experiments. The results of cell proliferative inhibition and clonogenic assay showed that SF enhanced the radiosensitivity of A549 cells when applied before 60Co γ-irradiation and this effect was mainly time and concentration dependent. The flow cytometric data indicated that SF treatment before the irradiation increased the G2/M phase, thus improving the radiosensitivity of A549, leading to cell apoptosis. This paper is the first study that describes the in vitro radiosensitising, cell cycle and apoptotic-inducing effects of schinifoline.

  3. Chemosensitivity of radioresistant cells in the multicellular spheroids of A549 lung adenocarcinoma

    Huang Gang

    2009-06-01

    Full Text Available Abstract Background The relapse of cancer after radiotherapy is a clinical knotty problem. Previous studies have demonstrated that the elevation of several factors is likely in some way to lead to the development of treatment tolerance, so it is necessary to further explore the problem of re-proliferated radioresistant cells to chemotherapeutic agents. In the present study, we aimed to investigate the chemosensitivity of radioresistant cells originated from the multicellular spheroids of A549 lung adenocarcinoma. Methods After irradiated with 25 Gy of 6 MV X-ray to A549 multicellular spheroids, whose 10th re-proliferated generations were employed as radioresistant cells, and the control groups were A549 parental cells and MCF7/VCR resistant cells. The chemo-sensitivity test was made by six kinds of chemotherapeutic drugs which were DDP, VDS, 5-Fu, HCP, MMC and ADM respectively, while verapamil (VPL was used as the reversal agent. Then the treatment effect was evaluated by MTT assay, and the multidrug resistant gene expressions of mdr1 and MRP were measured by RT-PCR. Results Both A549 parental cells and A549 derived radioresistant cells were resistant to DDP, but sensitive to VDS, 5-Fu, HCP, MMC and ADM. The inhibitory rates of VPL to these two types of cell were 98% and 25% respectively (P Mdr1/β2-MG and MRP/β2-MG of all A549 cells were about 0 and 0.7 respectively, and those of MCF7/VCR cells were 35 and 4.36. Conclusion The chemosensitivity of A549 radioresistant cells had not changed markedly, and the decreased sensitivity to VPL could not be explained by the gene expression of mdr1 and MRP. It is possible that the changes in the cell membrane and decreased proliferate ability might be attributed to the resistance. Unlike multidrug resistance induced by chemotherapy, VPL may be not an ideal reverser to radioresistant cells. Therefore, the new biological strategy needs to be developed to treat recurring radioresistant tumor in combination

  4. Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway

    Zhang, Jian, E-mail: zhangjian197011@yahoo.com [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi' an 710032 (China); Zhang, Tao [Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi' an 710038 (China); Ti, Xinyu; Shi, Jieran; Wu, Changgui; Ren, Xinling [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi' an 710032 (China); Yin, Hong, E-mail: yinnhong@yahoo.com [The Medical Image Center, Xijing Hospital, The Fourth Military Medical University, Xi' an 710032 (China)

    2010-08-13

    Research highlights: {yields} Curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells {yields} Curcumin promotes apoptosis in A549/DDP cells through a miRNA signaling pathway {yields} Curcumin induces A549/DDP cell apoptosis by downregulating miR-186* {yields} miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin -- Abstract: Curcumin extracted from the rhizomes of Curcuma longa L. has been shown to have inhibitory effects on cancers through its anti-proliferative and pro-apoptotic activities. Emerging evidence demonstrates that curcumin can overcome drug resistance to classical chemotherapies. Thus, the mechanisms underlying the anti-tumor activities of curcumin require further study. In our study, we first demonstrated that curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells. Further studies showed that curcumin altered miRNA expression; in particular, significantly downregulated the expression of miR-186* in A549/DDP. In addition, transfection of cells with a miR-186* inhibitor promoted A549/DDP apoptosis, and overexpression of miR-186* significantly inhibited curcumin-induced apoptosis in A549/DDP cells. These observations suggest that miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin.

  5. Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway

    Research highlights: → Curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells → Curcumin promotes apoptosis in A549/DDP cells through a miRNA signaling pathway → Curcumin induces A549/DDP cell apoptosis by downregulating miR-186* → miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin -- Abstract: Curcumin extracted from the rhizomes of Curcuma longa L. has been shown to have inhibitory effects on cancers through its anti-proliferative and pro-apoptotic activities. Emerging evidence demonstrates that curcumin can overcome drug resistance to classical chemotherapies. Thus, the mechanisms underlying the anti-tumor activities of curcumin require further study. In our study, we first demonstrated that curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells. Further studies showed that curcumin altered miRNA expression; in particular, significantly downregulated the expression of miR-186* in A549/DDP. In addition, transfection of cells with a miR-186* inhibitor promoted A549/DDP apoptosis, and overexpression of miR-186* significantly inhibited curcumin-induced apoptosis in A549/DDP cells. These observations suggest that miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin.

  6. 异长春花碱逆转肺癌顺铂耐药A549/DDP细胞耐药性的作用和机制%The Effect and Mechanism of Vinorelbine on Cisplatin Resistance of Human Lung Cancer Cell Line A549/DDP

    齐春胜; 高森; 李会强; 高卫真

    2014-01-01

    背景与目的肺癌细胞耐药已经成为肺癌化疗的主要困难之一,异长春花碱被认为可有效抑制肺癌细胞的增殖和转移。本研究旨在探讨异长春花碱对人肺癌A549/DDP细胞顺铂耐受性的逆转作用及机制。方法1μmol/L和5μmol/L异长春花碱作用A549/DDP细胞后,应用MTS法检测肿瘤细胞顺铂敏感性的变化,应用流式细胞术检测肿瘤细胞凋亡率变化,肿瘤细胞对Rh-123摄入量的变化,Western blot法检测MDR1、Bcl-2、survivin、caspase-3/8和PTEN蛋白表达以及Akt的磷酸化水平的变化,real-time PCR检测MDR1、Bcl-2、survivin和PTEN的mRNA表达,用报告基因系统检测NF-κB、Twist和Snail的转录活性。结果1μmol/L和5μmol/L异长春花碱作用A549/DDP细胞后,肿瘤细胞对顺铂的敏感性分别提高了1.91倍和2.54倍,肿瘤细胞对Rh-123的摄入量提高了1.93倍和2.95倍,细胞凋亡增加了2.25倍和3.82倍,MDR1、Bcl-2、survivin蛋白表达和Akt磷酸化水平下调,caspase-3/8和PTEN蛋白表达上调,MDR1的mRNA表达下调43.5%和25.8%,Bcl-2的mRNA表达下调57.3%和34.1%,survivin的mRNA表达下调37.6%和12.4%,PTEN表达上调183.4%和154.2%,NF-κB转录活性下降53.2%和34.5%,Twist转录活性下降61.4%和33.5%, Snail转录活性下降57.8%和18.7%。结论异长春花碱可提高肿瘤细胞A549/DDP对顺铂的敏感性,其机制可能与调节PTEN/AKT/NF-κB信号路径活性,进而下调耐药基因表达,上调促凋亡基因表达有关。%Background and objective Drug resistance is a major obstacle on lung cancer treatment and Vinorel-bine is an effective drug to inhibition of tumor proliferation and metastasis. In this study, we investigated the effect and mecha-nism of Vinorelbine on reversing the cisplatin resistance of human lung cancer A549/DDP cell line. Methods With 1μmol/L and 5μmol/L Vinorelbine treatment, MTS assay was employed to determine the effect of the cisplatin

  7. Cyclin Y和Cyclin X在肺癌细胞株A549中的细胞定位和功能%The Function Study and Cell Localization of Cyclin Y and Cyclin X in Lung Cancer Cell Line A549

    周世杰; 江姝; 赵晓婷; 岳文涛

    2013-01-01

    [Purpose] To construct pEGFP-N1/CCNY vector and pEGFP-N1/CCNX eukaryotic expression vector,and to explore the location and function of CyclinY/CyclinX in lung caner A549 cell.[Methods] CCNY and CCNX genes were amplified from human lung adenocarcinoma cell line H1299 by PCR.The recombinant plasmids pEGFP-N1/CCNY and pEGFP-N1/CCNX were constructed and transfected into A549 cells.The cellular localization and expression of CyclinY and Cyclin X were detected by fluorescence microscopy and Western Blot.[Results] The recombinant plasmid pEGFP-N1/CCNY and pEGFP-N1/CCNX were constructed successfully.Green fluorescence on the surface of transfected cells was found by fluorescence microscope.Western Blot confirmed Cyclin Y,Cyclin X expression.Cyclin Y and Cyclin X located at cellular membrane and nucleus in recombinant plasmid cell respectively.After transfection,A549-CCNY pEGFPN1 cell viability was 1.36±0.02,A549-CCNX pEGFPN cell viability was 11.45 ±0.05,which was higher than that in A549-pEGFPN1 (1.31±0.03) (P all<0.01).[Conclusion] In A549 cell,Cyclin Y and Cyclin X are differently distributed,Cyclin X plays more important role in promoting proliferation than Cyclin Y.%[目的]构建CCNY和CCNX基因的真核表达载体并观察其在人肺癌细胞株A549中的表达及定位,为进一步探讨Cyclin Y、Cyclin X在肺癌中的细胞定位和功能奠定了基础.[方法]以人肺腺癌细胞株H1299 cDNA为模板扩增CCNY和CCNX基因,并构建CCNY和CCNX过表达真核表达载体.应用荧光显微照相及Western Blot方法鉴定该细胞株中Cyclin Y、Cyclin X的定位及表达.[结果]成功构建pEGFP-N1/CCNY和pEGFP-N1/CCNX真核表达载体.荧光显微照相显示绿色荧光,Western Blot检测证实转染重组质粒细胞表达Cyclin Y、Cyclin X蛋白,Cyclin Y和Cyclin X分别定位于胞膜与胞核.A549-pEGFPN1细胞活性为1.31±0.03,而转染后的A549-CCNY pEGFPN1细胞活性为1.36±0.02,A549-CCNX pEGFPN1细胞活性为1.45±0.05(P<0

  8. Direct and in vitro observation of growth hormone receptor molecules in A549 human lung epithelial cells by nanodiamond labeling

    Cheng, C.-Y.; Perevedentseva, E.; Tu, J.-S.; Chung, P.-H.; Cheng, C.-L.; Liu, K.-K.; Chao, J.-I.; Chen, P.-H.; Chang, C.-C.

    2007-04-01

    This letter presents direct observation of growth hormone receptor in one single cancer cell using nanodiamond-growth hormone complex as a specific probe. The interaction of surface growth hormone receptor of A549 human lung epithelial cells with growth hormone was observed using nanodiamond's unique spectroscopic signal via confocal Raman mapping. The growth hormone molecules were covalent conjugated to 100nm diameter carboxylated nanodiamonds, which can be recognized specifically by the growth hormone receptors of A549 cell. The Raman spectroscopic signal of diamond provides direct and in vitro observation of growth hormone receptors in physiology condition in a single cell level.

  9. Interleukin 23 Promotes Lung Adenocarcinoma A549 Cell Migration and Invasion

    Sen ZHANG

    2012-05-01

    Full Text Available Background and objective Interleukin 23 (IL-23 is a pro-inflammatory cytokine that plays an important role in inflammatory disease and tumor microenvironment. The IL-23 receptor is expressed in colon, lung, and oral carcinomas. We performed this study to investigate whether IL-23 promotes directly carcinoma cell migration and invasion as well as further explore its mechanism. Methods The migration and invasion of human lung adenocarcinoma A549 cells induced by IL-23 were detected by a scratch test and Transwell experiment. MMP-9 expression of the mRNA and protein levels of A549 cells cultured with and without IL-23 was respectively detected by Real-time PCR and ELISA. The effect of IL-23 on A549 cells was further verified using anti-IL-23p19 neutralization antibody (Ab IL-23p19 to eliminate IL-23. Results IL-23 remarkably promoted A549 cell migration and invasion. MMP-9 expression in A549 cells was upregulated by IL-23 stimulation. In addition, the effect of IL-23 on the migration and invasion of A549, as well as the MMP-9 expression in A549 cells induced by IL-23, was eliminated by Ab IL-23p19. Conclusion IL-23 promotes the migration and invasion of A549 cells by inducing MMP-9 expression.

  10. Effect of staurosporine on the mobility and invasiveness of lung adenocarcinoma A549 cells: an in vitro study

    Lung cancer is one of the most malignant tumors, representing a significant threat to human health. Lung cancer patients often exhibit tumor cell invasion and metastasis before diagnosis which often render current treatments ineffective. Here, we investigated the effect of staurosporine, a potent protein kinase C (PKC) inhibitor on the mobility and invasiveness of human lung adenocarcinoma A549 cells. All experiments were conducted using human lung adenocarcinoma A549 cells that were either untreated or treated with 1 nmol/L, 10 nmol/L, or 100 nmol/L staurosporine. Electron microscopy analyses were performed to study ultrastructural differences between untreated A549 cells and A549 cells treated with staurosporine. The effect of staurosporine on the mobility and invasiveness of A549 was tested using Transwell chambers. Western blot analyses were performed to study the effect of staurosporine on the levels of PKC-α, integrin β1, E-cadherin, and LnR. Changes in MMP-9 and uPA levels were identified by fluorescence microscopy. We demonstrated that treatment of A549 cells with staurosporine caused alterations in the cell shape and morphology. Untreated cells were primarily short spindle- and triangle-shaped in contrast to staurosporine treated cells which were retracted and round-shaped. The latter showed signs of apoptosis, including vacuole fragmentation, chromatin degeneration, and a decrease in the number of microvilli at the surface of the cells. The A549 cell adhesion, mobility, and invasiveness significantly decreased with higher staurosporine concentrations. E-cadherin, integrin β1, and LnR levels changed by a factor of 1.5, 0.74, and 0.73, respectively compared to untreated cells. In addition, the levels of MMP-9 and uPA decreased in cells treated with staurosporine. In summary, this study demonstrates that staurosporine inhibits cell adhesion, mobility, and invasion of A549 cells. The staurosporine-mediated inhibition of PKC-α, induction of E

  11. E2F1 enhances 8-chloro-adenosine-induced G2/M arrest and apoptosis in A549 and H1299 lung cancer cells.

    Duan, Hong-Ying; Cao, Ji-Xiang; Qi, Jun-Juan; Wu, Guo-Sheng; Li, Shu-Yan; An, Guo-Shun; Jia, Hong-Ti; Cai, Wang-Wei; Ni, Ju-Hua

    2012-03-01

    The E2F1 transcription factor is a well known regulator of cell proliferation and apoptosis, but its role in response to DNA damage is less clear. 8-Chloro-adenosine (8-Cl-Ado), a nucleoside analog, can inhibit proliferation in a variety of human tumor cells. However, it is still elusive how the agent acts on tumors. Here we show that A549 and H1299 cells formed DNA double-strand breaks after 8-Cl-Ado exposure, accompanied by E2F1 upregulation at protein level. Overexpressed wild-type (E2F1-wt) colocalized with double-strand break marker γ-H2AX and promoted G2/M arrest in 8-Cl-Ado-exposed A549 and H1299, while expressed S31A mutant of E2F1 (E2F1-mu) significantly reduced ability to accumulate at sites of DNA damage and G2/M arrest, suggesting that E2F1 is required for activating G2/M checkpoint pathway upon DNA damage. Transfection of either E2F1-wt or E2F1-mu plasmid promoted apoptosis in 8-Cl-Ado-exposed cells, indicating that 8-Cl-Ado may induce apoptosis in E2F1-dependent and E2F1-independent ways. These findings demonstrate that E2F1 plays a crucial role in 8-Cl-Ado-induced G2/M arrest but is dispensable for 8-Cl-Ado-induced apoptosis. These data also suggest that the mechanism of 8-Cl-Ado action is complicated. PMID:22803943

  12. 5-allyl-7-gen-difluoromethoxychrysin enhances TRAIL-induced apoptosis in human lung carcinoma A549 cells

    5-allyl-7-gen-difluoromethoxychrysin (AFMC) is a novel synthetic analogue of chrysin that has been reported to inhibit proliferation in various cancer cell lines. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. The cytotoxicity of A549 and WI-38 cells were determined using colorimetry. Apoptosis was detected by flow cytometry (FCM) after propidium iodide (PI) fluorescence staining and agarose gel electrophoresis. Caspase activities were evaluated using enzyme-linked immunosorbent assay (ELISA).The expressions of DR4 and DR5 were analyzed using FCM and western blot. Subtoxic concentrations of AFMC sensitize human non-small cell lung cancer (NSCLC) A549 cells to TRAIL-mediated apoptosis. Combined treatment of A549 cells with AFMC and TRAIL significantly activated caspase-3, -8 and -9. The caspase-3 inhibitor zDEVD-fmk and the caspase-8 inhibitor zIETD-fmk blocked the apoptosis of A549 cells induced by co-treatment with AFMC and TRAIL. In addition, we found that treatment of A549 cells with AFMC significantly induced the expression of death receptor 5 (DR5). AFMC-mediated sensitization of A549 cells to TRAIL was efficiently reduced by administration of a blocking antibody or small interfering RNAs against DR5. AFMC also caused increase of the Sub-G1 cells by TRAIL treatment and increased the expression levels of DR5 in other NSCLC H460 and H157 cell lines. In contrast, AFMC-mediated induction of DR5 expression was not observed in human embryo lung WI-38 cells, and AFMC did not sensitize WI-38 cells to TRAIL-induced apoptosis. AFMC synergistically enhances TRAIL-mediated apoptosis in NSCLC cells through up-regulating DR5 expression

  13. A preliminary study on radiosensitization effect of curcumin plus cisplatin on non-small cell lung cancer A549 cells%姜黄素联合顺铂对非小细胞肺癌细胞A549放疗增敏作用的初步研究

    蔡勇; 王季颖

    2015-01-01

    Objective To explore the radiosensitization effect of curcumin plus cisplatin on non⁃small cell lung cancer A549 cells. Methods Cell viability at 24, 48 and 72 h after treatment with different concentrations ( 10, 20, 50, 100, 200 μmol/L) of curcumin or ( 1, 2, 5, 10, 20 mg/L) of cisplatin were determined by MTT. According to the experimental protocol, the below experi⁃ments were carried out in irradiation ( R ) group, curcumin+irradiation ( C+R ) group, cisplatin+irradiation ( P+R ) group and curcumin+cisplatin+irradiation ( C+P+R) group. The colony formation assay was employed to observe the surviving fraction ( SF) of a⁃bove four groups after X⁃ray irradiation of 0, 2, 4, 6, 8, 10 Gy. The artificial scratch, Transwell test and Western blotting were em⁃ployed to detect the cell migration, invasion and protein level of epidermal growth factor receptor ( EGFR) at 24 h after treatment in four groups. Results The cell viability of A549 cells gradually decreased with the increasing concentration of curcumin ranging from 10 to 200 μmol/L and cisplatin ranging from 1 to 20 mg/L in a dose⁃and time⁃dependent manner ( P<0�05) . The SF of C+P+R group were lower than the remaining 3 groups under the dose of 2⁃10 Gy ( P<0�05) . Compared with the R irradiation group, there was lower SF in C+R group under the dose of 4⁃10 Gy and P+R group under the dose of 2⁃10 Gy with significant difference ( P<0�05) . Compared with R group, the sensitizing enhancement ratio were 1�24, 1�31 and 1�96 in C+R group, P+R group and C+P+R group, respective⁃ly. There were lower migration distance, transmembrane cell number and EGFR protein level in C+P+R group versus the remaining 3 groups ( P<0�05) . Compared with the R group, the above indicators were also lower in C+R group and P+R group under the dose of 2⁃10 Gy with significant difference ( P<0�05) . Conclusion Curcumin plus cisplatin can inhibit the proliferation of A549 cells with radi

  14. 人肺腺癌A549细胞低剂量辐射超敏感性及其机制的研究%Low dose hyper-radiosensitivity in human lung cancer cell line A549 and its possible mechanisms

    陶丹; 程晶; 伍钢; 吴红革; 薛军

    2009-01-01

    目的 观察A549细胞的低剂量辐射超敏感性现象,探讨其发生的机制.方法 A549细胞接受0~2 Gy的60Co γ射线照射后,流式细胞仪对其分选计数,克隆形成法检测细胞存活分数,Western blot法检测ATMl981Ser-P蛋白表达,Hoechst 33258荧光染色法、AnnexinV-FITC/PI双染流式细胞仪检测细胞凋亡,PI单染流式细胞仪检测细胞周期.结果 细胞在0~0.3 Gy表现出单位剂量杀伤增强,在0.3~0.5 Gy表现出一定的辐射抗性,0.5 Gy后的区域存活分数随辐射剂量的增加而降低.照射后1 h,ATM激酶在0.2 Gy时开始活化,0.5 Gy时活化达高峰(t=7.96,P<0.05);与0.5 Gy相比1.0和2.0 Gy的活化水平无明显变化(t=0.69、0.55,P>0.05).照射后24 h,部分细胞发生凋亡,其凋亡曲线与存活曲线相吻合.与未照射组相比,0.1和0.2 Gy组在各时间点(照射后6、12和24 h)的细胞周期无明显变化,而0.3、0.4和0.5 Gy组,照射后6和12 h细胞发生G2/M期阻滞(t=2.87、2.88、4.92和3.70、3.12、8.11,P<0.05),照射后24 h G2/M期细胞比例下降(t=3.87、4.77、3.01,P<0.05).结论 A549细胞存在HRS/IRR现象,其发生可能与ATM激酶、细胞周期变化有关,凋亡是细胞死亡的主要方式.%Objective To study the low dose hyper-radiosensitivity in human lung cancer cell line A549,and its possible mechanisms.Methods Exponentially growing A549 cells were irradiated with 60Co γ-rays at doses of 0-2 Gy.Together with flow cytometry for precise cell sorting,cell survival fraction was measured by mean of conventional colony-formation assay.ATM1981 Ser-P protein expression was examined by Western blot.Apoptosis was identified by Hoechst 33258 fluorescent staining,and Annexin V-FITC and propidium iodide staining flow cytometry.Cell cycle distribution was observed by flow cytometry.Results There was an excessive cell killing per unit dose when the doses were below about 0.3 Gy,and the cells exhibited more resistant response at the doses between

  15. High throughput determination of TGFβ1/SMAD3 targets in A549 lung epithelial cells.

    Yingze Zhang

    Full Text Available BACKGROUND: Transforming growth factor beta 1 (TGFβ1 plays a major role in many lung diseases including lung cancer, pulmonary hypertension, and pulmonary fibrosis. TGFβ1 activates a signal transduction cascade that results in the transcriptional regulation of genes in the nucleus, primarily through the DNA-binding transcription factor SMAD3. The objective of this study is to identify genome-wide scale map of SMAD3 binding targets and the molecular pathways and networks affected by the TGFβ1/SMAD3 signaling in lung epithelial cells. METHODOLOGY: We combined chromatin immunoprecipitation with human promoter region microarrays (ChIP-on-chip along with gene expression microarrays to study global transcriptional regulation of the TGFβ1/SMAD3 pathway in human A549 alveolar epithelial cells. The molecular pathways and networks associated with TGFβ1/SMAD3 signaling were identified using computational approaches. Validation of selected target gene expression and direct binding of SMAD3 to promoters were performed by quantitative real time RT-PCR and electrophoretic mobility shift assay on A549 and human primary lung epithelial cells. RESULTS AND CONCLUSIONS: Known TGFβ1 target genes such as SERPINE1, SMAD6, SMAD7, TGFB1 and LTBP3, were found in both ChIP-on-chip and gene expression analyses as well as some previously unrecognized targets such as FOXA2. SMAD3 binding of FOXA2 promoter and changed expression were confirmed. Computational approaches combining ChIP-on-chip and gene expression microarray revealed multiple target molecular pathways affected by the TGFβ1/SMAD3 signaling. Identification of global targets and molecular pathways and networks associated with TGFβ1/SMAD3 signaling allow for a better understanding of the mechanisms that determine epithelial cell phenotypes in fibrogenesis and carcinogenesis as does the discovery of the direct effect of TGFβ1 on FOXA2.

  16. Irradiation and various cytotoxic drugs enhance tyrosine phosphorylation and {beta}{sub 1}-integrin clustering in human A549 lung cancer cells in a substratum-dependent manner in vitro

    Cordes, N.; Beinke, C.; Beuningen, D. van [Inst. of Radiobiology, German Armed Forces, Munich (Germany); Plasswilm, L. [Dept. of Radiation Oncology, Univ. Hospital Basel (Swaziland)

    2004-03-01

    Background and purpose: interactions of cells with a substratum, especially extracellular matrix proteins, initiate clustering of integrin receptors in the cell membrane. This process represents the initial step for the activation of signaling pathways regulating survival, proliferation, differentiation, adhesion, and migration, and could, furthermore, be important for cellular resistance-mediating mechanisms against radiation or cytotoxic drugs. The lack of data elucidating the impact of irradiation or cytotoxic drugs on this important phenomenon led to this study on human A549 lung cancer cells in vitro. Material and methods: the human lung carcinoma cell line A549 grown on polystyrene or fibronectin (FN) was irradiated with 0-8 Gy or treated with cisplatin (0.1-50 {mu}M), paclitaxel (0.1-50 nM), or mitomycin (0.1-50 {mu}M). Colony formation assays, immunofluorescence staining in combination with activation of integrin clustering using anti-{beta}{sub 1}-integrin antibodies (K20), and Western blotting for tyrosine phosphorylation under treatment of cells with the IC{sub 50} for irradiation (2 Gy; IC{sub 50} = 2.2 Gy), cisplatin (2 {mu}M), paclitaxel (5 nM), or mitomycin (7 {mu}M) were performed. Results: attachment of cells to FN resulted in a significantly reduced radio- and chemosensitivity compared to polystyrene. The clustering of {beta}{sub 1}-integrins examined by immunofluorescence staining was only stimulated by irradiation, cisplatin, paclitaxel, or mitomycin in case of cell attachment to FN. By contrast, tyrosine phosphorylation, as one of the major events following {beta}{sub 1}-integrin clustering, showed a 3.7-fold, FN-related enhancement, and treatment of cells with the IC{sub 50} of radiation, cisplatin, paclitaxel, or mitomycin showed a substratum-dependent induction. Conclusion: for the first time, a strong influence of irradiation and a variety of cytotoxic drugs on the clustering of {beta}{sub 1}-integrins could be shown. This event is a

  17. Inhibition of mitogen activated protein kinases increases the sensitivity of A549 lung cancer cells to the cytotoxicity induced by a kava chalcone analog

    Janel K Warmka; Solberg, Eric L.; Zeliadt, Nicholette A.; Srinivasan, Balasubramanian; Charlson, Aaron T.; Xing, Chengguo; Wattenberg, Elizabeth V.

    2012-01-01

    We are interested in investigating the biological activity of chalcones, a major class of compounds found in the beverage kava, in order to develop potent and selective chemopreventive candidates. Consumption of kava in the South Pacific Islands is inversely correlated with cancer incidence, even among smokers. Accordingly, chalcones have anti-cancer activities in animal and cell culture models. To investigate signaling pathways that affect chalcone action we studied a potent analog, (E)-3-(3...

  18. 去甲斑蝥素对人肺腺癌A549细胞的抑制作用%The inhibition of norcantharidin on human lung adenocarcinoma A549 cells

    崔宝弟; 王敏; 孙震晓

    2015-01-01

    OBJECTIVE:To study the effects of norcantharidin (NCTD) on human lung cancer cells,and investigate the mechanisms.METHODS:The growth inhibition of A549 cells treated with 0-240µmol/L NCTD for 0-72 hours was analyzed by MTT assay. The recovery growth and proliferation of A549 cells treated with 0-120µmol/L NCTD for 24 h was evaluated by MTT assay. The morphological changes of A549 cells treated with 40,50 and 60µmol/L NCTD for 0-72 h were examined under inverted microscope. The apoptosis and cell cycle changes of A549 cells treated with 40-60µmol/L NCTD were detected by flow cytometry.RESULTS:NCTD inhibited the growth of A549 cells in 30-240 µmol/L(P0.05)。结论:40~60µmol/L NCTD主要通过诱导A549细胞G2~M期阻滞而抑制细胞生长。

  19. Research of vaccination with whole cells antigens from mesenchymal stem cells generate an antitumor effect of lung cancer cell line A549 in vivo%骨髓间充质干细胞全细胞抗原干预肺癌细胞系A549移植瘤生长的实验研究

    李静; 陈军; 李秀玉

    2014-01-01

    目的:观察骨髓间充质干细胞(MSCs)的全细胞抗原(WCAs)对人肺腺癌细胞株A549荷瘤的影响,并探讨肿瘤相关增殖抗原的改变揭示其可能的抑制机制。方法全骨髓贴壁法原代培养小鼠MSCs,取3~5代MSCs以15 Gy X线灭活获取WCAs,将BALB/c小鼠随机分成实验组和对照组,每组各24只。实验组皮下接种WCAs(1次/3d,共2周),获得免疫接种小鼠模型,对照组皮下注射同体积的磷酸缓冲液。观察两组小鼠肿瘤生长情况,测量肿瘤直径、计算肿瘤体积,并于接种后第7(Day7)、30天(Day30)行Western blot及实时荧光定量逆转录聚合酶链反应检测增殖细胞核抗原(PCNA)及Ki-67因子的蛋白及mRNA水平。结果肺腺癌A549细胞皮下移植成功使小鼠荷瘤,实验组小鼠的肿瘤体积显著小于对照组(P<0.05);实验组PCNA蛋白水平显著低于对照组[Day7:(6.42±0.54)比(18.67±0.96),P<0.01;Day30:(2.12±0.14)比(4.32±0.25),P<0.05];PCNA mRNA水平低于对照组[Day7:(11.64±0.28)比(25.18±1.37),P<0.01;Day30:(2.11±0.18)比5.69±0.41),P<0.01];实验组Ki-67蛋白水平显著低于对照组[Day7:(1.57±0.51)比(4.84±0.23),P<0.05;Day30:(2.75±0.28)比(5.66±0.19),P<0.01];Ki-67 mRNA水平也明显下调[Day7:(2.12±0.43)比(5.94±1.03),P<0.01;Day30:(3.71±0.72)比(8.62±0.35),P<0.01]。结论采用MSCs获得全细胞抗原进行免疫应激可产生抑制肿瘤生长作用,其机制可能与及肿瘤增殖相关因子PCNA、Ki-67的下调有关,其内在的免疫分子机制有待深层次的探索。%Objective To observe the influence of whole cell antigens (WCAs) of mesenchymal stem cells (MSCs) on lung cancer cell line A549, discuss the change of related antigen of tumor proliferation, and reveal the possible inhibition mechanism. Methods The MSCs was isolated and cultured adherent cells from marrow, and then 3-5 generations MSCs were inactivated by X-ray (15 Gy), and the WCAs were obtained the BALB/c rats

  20. Hypoxia Upregulates the Expression of Annexin A1 in Lung Adenocarcinoma A549 Cells

    Zhenhong HU

    2012-05-01

    Full Text Available Background and objective The growth of tumor often faced up with lackness of blood and oxygen, and it has been reported that Annexin A1 may be involved in tumor. The aim of this investigation is to explore the characteristics of expression of Annexin A1 in lung adenocarcinoma A549 cells after hypoxia. Methods A549 cells were exposured to either normoxia (21%O2 or hypoxia (1%O2 condition for 4 h, 12 h, 24 h. The expressions of Annexin A1 mRNA levels were measured by RT-PCR. The expressions of Annexin 1 protein were investigaged by Western blot. The relative content of reactive oxygen species (ROS were assayed by special kit. The expressions of nuclear translocation of NF-κB was assayed by Western blot; After been treated with ROS scavenger NAC and PDTC, the levels of Annexin 1 protein of A549 cells were measured by Western blot. Results Compared with normoxia group, the Annexin A1 mRNA in hypoxia group increased after 4 h, and then decreased gradually; Moreover, Annexin 1 protein levels of A549 cells were also increased when treated with hypoxia. An increaing of ROS production in cells exprosed to hypoxia was detected. NAC and PDTC inhibited hypoxia-induced Annexin A1 increase. Conclusion Hypoxia upregulates the expression of Annexin A1 in lung adenocarcinoma A549 cells, in which process ROS-NF-κB may paticipate in.

  1. Transcription Activity of Ectogenic Human Carcinoembryonic Antigen Promoter in Lung Adenocarcinoma Cells A549

    XIONG Weining; FANG Huijuan; XU Yongjian; XIONG Shendao; CAO Yong; SONG Qingfeng; ZENG Daxiong; ZHANG Huilan

    2006-01-01

    The transcription activity of ectogenic human carcinoembryonic antigen (CEA) promoter in lung adenocarcinoma cells A549 was investigated for the further gene-targeting therapy. The reporter gene green fluorescent protein (GFP) driven by CEA promoter and human cytomegalovirus (CMV) promoter were relatively constructed and named plasmid pCEA-EGFP and pCMV-GFP respectively. The intensity of fluorescence was detected by fluorescence microscope and flow cytometry analysis after the pCEA-GFP and pSNAV-GFP plasmids were transfected into A549 cells through liposome respectively. The results showed (4.08±0.63) % of the A549 cells transfected with pCEA-AFP plasmid expressed, significantly lower than that of the A549 cells transfected with pCMV-GFP [(43.27±3.54) %]. It was suggested that ectogenic human CEA promoter in lung adenocarcinoma cells A549 was weakly expressed. The distinct specificity of CEA promoter in CEA high expression cells was regarded as a tool in selective gene therapy, but the transcription activity of ectogenic human CEA promoter was needed to increase in the future.

  2. Nimesulide has a role of radio-sensitizer against lung carcinoma A549 cells

    Won, Joo Yoon; Park, Jong Kuk; Hong, Sung Hee [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2006-07-01

    Cyclooxygenases (COX) are key enzymes in the prostaglandin synthesis. There are two isoforms of the COX enzyme, COX-1 and COX-2. COX-2 expression is associated with carcinogenesis in variety of cancers and to render cells resistant to apoptotic stimuli. Increased expression of COX-2 is shown in non-small cell lung cancer (NSCLC), specifically in adenocarcinomas. Radiotherapy has been the important treatment for NSCLC. In recent studies, newer molecules that target specific pathophysiology or molecular pathways have been tested for the radiation sensitizers. COX-2 inhibitors are shown to enhanced radioresponse of cultured human cancer cell lines and immunodeficient mice. However, little is known about the molecular and biochemical mechanisms how NSAIDs enhance radioresponse of tumor cells. Nimesulide (methanesulfonamide, N-(4-nitro-2- phenoxyphenyl)), selective COX-2 inhibitors, is a drug with anti-inflammatory, anti-pyretic and analgesic properties. Nimesulide has the specific affinity to inhibit the inducible form of cyclooxygenase (COX-2) rather than the constitutive form (COX-1), and is well tolerated by adult, elderly and pediatric patients. Nimesulide was found also to have a chemopreventive activity against colon, urinary bladder, breast, tongue, and liver carcinogenesis. In this study, we examined whether nimesulide can increase radiation induced cell death and its mechanism in NSCLC cells A549.

  3. Nimesulide has a role of radio-sensitizer against lung carcinoma A549 cells

    Cyclooxygenases (COX) are key enzymes in the prostaglandin synthesis. There are two isoforms of the COX enzyme, COX-1 and COX-2. COX-2 expression is associated with carcinogenesis in variety of cancers and to render cells resistant to apoptotic stimuli. Increased expression of COX-2 is shown in non-small cell lung cancer (NSCLC), specifically in adenocarcinomas. Radiotherapy has been the important treatment for NSCLC. In recent studies, newer molecules that target specific pathophysiology or molecular pathways have been tested for the radiation sensitizers. COX-2 inhibitors are shown to enhanced radioresponse of cultured human cancer cell lines and immunodeficient mice. However, little is known about the molecular and biochemical mechanisms how NSAIDs enhance radioresponse of tumor cells. Nimesulide (methanesulfonamide, N-(4-nitro-2- phenoxyphenyl)), selective COX-2 inhibitors, is a drug with anti-inflammatory, anti-pyretic and analgesic properties. Nimesulide has the specific affinity to inhibit the inducible form of cyclooxygenase (COX-2) rather than the constitutive form (COX-1), and is well tolerated by adult, elderly and pediatric patients. Nimesulide was found also to have a chemopreventive activity against colon, urinary bladder, breast, tongue, and liver carcinogenesis. In this study, we examined whether nimesulide can increase radiation induced cell death and its mechanism in NSCLC cells A549

  4. Effects of Sodium Cantharidate Vitamin B6 on Proliferation,Apoptosis and Influence of NF-κB and Caspase3/7 on Human Lung Cancer A549 Cells%斑蝥酸钠维生素B6注射液对人肺癌细胞系A549增殖抑制及核因子κB和Caspase3/7的影响

    温省初; 王一飞; 李爱明; 李冠军; 成志勇; 王亚丽; 石林

    2011-01-01

    Objective To investigate the effect of sodium cantharidinate ( SC ) vitamin B6 on human non - small cell lung cancer A549 cell proliferation, apoptosis and the influence of transcription factor NF - kB and apoptosis molecules Caspase3/7. Methods Different concentrations of SC vitamin B6 and A549 cells were cultured together; Cells apoptosis was tested by light microscopy and fluorescent staining Hoechst33342 morphology; MTT assay tested cell proliferation; Rhodamine 123 examined mitochondrial membrane potential; Caspase3/7 activity assay kit tested Caspase3/7 activity; Western blot detected of NF - kB P65 , I - kB protein levels. Results SC vitamin B6 inhibited the A549 cells proliferation, of which there were apparent apoptotic morphological changes. When 5. 0 mg/L group roled in A549 cells 72 h, cell proliferation inhibition rate reached 67. 37 percent maximum. Mitochondrial membrane potential results showed that with increasing concentration of SC vitamin B6 and time, the mitochondrial membrane potential gradually weakened, while Caspase3/7 protein activity increased. After SC vitamin B6 was added in A549 cells, NF - kB P65 protein levels was reduced ( P < 0. 05 ) and I - kB protein levels had no changes. Conclusion SC vitamin B6 inhibits the NF - kB P65 expression, activates caspase - 3/7 activities which inhibits A549 cells proliferation and induce apoptosis.%目的 探讨斑蝥酸钠(SC)维生素B6注射液对人非小细胞肺癌A549细胞增殖、凋亡及核因子κB(NF-κB)、凋亡分子Caspase3/7的影响.方法 用不同浓度(0、1.0、2.5、5.0 mg/L)的SC维生素B6注射液处理A549细胞,观察光镜及Hoechst33342荧光染色检测细胞凋亡形态;用噻唑蓝(MTT)比色法检测SC维生素B6注射液对细胞增殖的抑制作用;罗丹明123检测线粒体膜电位;Caspase3/7活性检测试剂盒检测Caspase3/7活性;蛋白印迹检测NF-κB P65、I-κB 蛋白表达.结果 SC维生素B6注射液对A549细胞的体外增殖有明显抑制作

  5. Antineoplastic effects of deoxyelephantopin,a sesquiterpene lactone from Elephantopus scaber, on lung adenocarcinoma (A549) cells

    Farha A.Kabeer; Geetha B.Sreedevi; Mangalam S.Nair; Dhanya S.Rajalekshmi; Latha P.Gopalakrishnan; Sujathan Kunjuraman; Remani Prathapan

    2013-01-01

    OBJECTIVE:Deoxyelephantopin,a sesquiterpene lactone from Elephantopus scaber,showed inhibition of the growth of various tumor cells in vitro.In the present study,we investigated the cytotoxicity and apoptosis-inducing capacity of deoxyelephantopin on lung adenocarcinoma (A549) cells.METHODS:The cytotoxic effect of deoxyelephantopin on A549 cells and normal lymphocytes was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 50% inhibitory concentration (IC50) value was determined.The self-renewal and proliferating potential of A549 cells after treatment with deoxyelephantopin were examined by colony formation assay.Cellular morphology of deoxyelephantopin-treated cells was observed using phasecontrast microscopy.The induction of apoptosis was evaluated using acridine orange and ethidium bromide staining,Hoechst 33342 staining,terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling (TUNEL) assay,DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate staining by flow cytometry.Activation of caspases was detected using fluorogenic substrate specific to caspases 2,3,8 and 9 and flow cytometric analysis.The total cellular DNA content and expression of cleaved poly (ADP-ribose) polymerase was also analyzed.RESULTS:Deoxyelephantopin exhibited cytotoxicity to A549 cells (IC5o =12.287 μg/mL),however,there was no toxicity towards normal human lymphocytes.Deoxyelephantopin suppressed the colony-forming ability of A549 cells in a dose-dependent manner.Acridine orange,ethidium bromide and Hoechst 33342 staining showed cell shrinkage,chromosomal condensation and nuclear fragmentation,indicating induction of apoptosis.Deoxyelephantopin increased apoptosis of A549 cells,as evidenced by more TUNEL-positive cells.DNA fragmentation and Annexin V staining revealed late-stage apoptotic cell population.Deoxyelephantopin inhibited A549 cell growth by cell cycle arrest at G2/M phase and induced apoptosis through

  6. Human lung epithelial cell A549 proteome data after treatment with titanium dioxide and carbon black.

    Vuong, Ngoc Q; Goegan, Patrick; Mohottalage, Susantha; Breznan, Dalibor; Ariganello, Marianne; Williams, Andrew; Elisma, Fred; Karthikeyan, Subramanian; Vincent, Renaud; Kumarathasan, Premkumari

    2016-09-01

    Here, we have described the dataset relevant to the A549 cellular proteome changes after exposure to either titanium dioxide or carbon black particles as compared to the non-exposed controls, "Proteomic changes in human lung epithelial cells (A549) in response to carbon black and titanium dioxide exposures" (Vuong et al., 2016) [1]. Detailed methodologies on the separation of cellular proteins by 2D-GE and the subsequent mass spectrometry analyses using MALDI-TOF-TOF-MS are documented. Particle exposure-specific protein expression changes were measured via 2D-GE spot volume analysis. Protein identification was done by querying mass spectrometry data against SwissProt and RefSeq protein databases using Mascot search engine. Two-way ANOVA analysis data provided information on statistically significant A549 protein expression changes associated with particle exposures. PMID:27508218

  7. Inhibitory Effect of Cantharidin on Proliferation of A549 Cells

    WANG Xiao-hua; YIN Yuan-qin; SUI Cheng-guang; MENG Fan-dong; MA Ping; JIANG You-hong

    2007-01-01

    Objective: To study the inhibition of Cantharidin against the proliferation of human lung cancer A549 cells and its mechanism. Methods: MTT assay was employed to determine the inhibition of Cantharidin against proliferation of A549 cells and flow Cytometry was applied to analyze A549 cell cycle and the effect of Cantharidin on cell cycle. Results: Cantharidin showed inhibition against the proliferation of A549 cells, and the inhibition was mediated by blocking A549 cell cycle at G2/M phase significantly. Conclusion: Cantharidin exhibits inhibition against the proliferation of human lung cancer A549 cells.

  8. Role of Rad52 in fractionated irradiation induced signaling in A549 lung adenocarcinoma cells

    Ghosh, Somnath, E-mail: ghosh.barc@gmail.com [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India); Krishna, Malini, E-mail: malinik00@gmail.com [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India)

    2012-01-03

    The effect of fractionated doses of {gamma}-irradiation (2 Gy per fraction over 5 days), as delivered in cancer radiotherapy, was compared with acute doses of 10 and 2 Gy, in A549 cells. A549 cells were found to be relatively more radioresistant if the 10 Gy dose was delivered as a fractionated regimen. Microarray analysis showed upregulation of DNA repair and cell cycle arrest genes in the cells exposed to fractionated irradiation. There was intense activation of DNA repair pathway-associated genes (DNA-PK, ATM, Rad52, MLH1 and BRCA1), efficient DNA repair and phospho-p53 was found to be translocated to the nucleus of A549 cells exposed to fractionated irradiation. MCF-7 cells responded differently in fractionated regimen. Silencing of the Rad52 gene in fractionated group of A549 cells made the cells radiosensitive. The above result indicated increased radioresistance in A549 cells due to the activation of Rad52 gene.

  9. Role of Rad52 in fractionated irradiation induced signaling in A549 lung adenocarcinoma cells.

    Ghosh, Somnath; Krishna, Malini

    2012-01-01

    The effect of fractionated doses of γ-irradiation (2Gy per fraction over 5 days), as delivered in cancer radiotherapy, was compared with acute doses of 10 and 2Gy, in A549 cells. A549 cells were found to be relatively more radioresistant if the 10Gy dose was delivered as a fractionated regimen. Microarray analysis showed upregulation of DNA repair and cell cycle arrest genes in the cells exposed to fractionated irradiation. There was intense activation of DNA repair pathway-associated genes (DNA-PK, ATM, Rad52, MLH1 and BRCA1), efficient DNA repair and phospho-p53 was found to be translocated to the nucleus of A549 cells exposed to fractionated irradiation. MCF-7 cells responded differently in fractionated regimen. Silencing of the Rad52 gene in fractionated group of A549 cells made the cells radiosensitive. The above result indicated increased radioresistance in A549 cells due to the activation of Rad52 gene. PMID:22001234

  10. Chemosensitivity of irradiated resistant cells of multicellular spheroids in A549 lung adenocarcinoma

    Objective: To investigate the chemosensitivity of irradiated resistant cells of multicellular spheroids in A549 lung adenocarcinoma. Methods: The A549 irradiated resistant cells were the 10th regrowth generations after irradiated with 2.5 Gy of 6 MV X-ray, the control groups were A549 parent cells and MCFY/VCR resistant cells. The 6 kinds of chemotherapeutic drugs were DDP, VDS, 5-FU, HCP, MMC and ADM respectively, with verapamil (VPL) as reverse agent. The treatment effect was compared with MTT assay, and the multidrug resistant gene expressions of mdrl and MRP were measured with RT-PCR method. Results: A549 cells and irradiated resistant cells were resistant to DDP, but sensitivity to VDS,5-FU, HCP, MMC and ADM. The inhibitory rates of VPL to the above two cells were 98% and 25% respectively(P2-MG and MRP/β2-MG of all A549 cells were about 0 and 0.7 respectively, and those of MCFT/VCR cells were 35 and 4.36. Conclusion: The chemosensitivity of A549 irradiated resistant cells had not changed markedly, the decreased sensitivity to VPL could not be explained by the gene expression of mdrl and MRP. It is conferred that some kinds of changes in the cell membrane and decreased regrowth ability to result in resistance. Unlike multidrug resistance induced by chemotherapy, VPL may be not an ideal reverser to irradiated resistant cells. The new kinds of biological preparation should be sought to combine chemotherapy to treat recurring tumor with irradiated resistance. (authors)

  11. Enrichment and identification of lung adenocarcinoma initiating cells from A 549%A549肺腺癌始动细胞的富集和鉴定

    林盛; 张振华; 饶明月; 吴敬波

    2013-01-01

    Objective To obtain the lung adenocarcinoma initiating cells from the A 549 cell line based on paclitaxel treatment combination with serum-free cultivation and to validate spared cells can represent tumor initiating cells (TICs) .Methods After dis-sociated by trypsogen ,about 106 /mL cells were suspended in serum-free medium supplemented with 0 .4% bovine serum albumin (BSA) ,insulin ,basic fibroblast growth factor (bFGF) ,human recombinant epidermal growth factor (EGF) and obtained spheroid cells .At the second passage ,paclitaxel was added at a concentration of 100 nmol/L for 48 h and then replaced with completely fresh medium once or twice per week until new spheroids emerged .Results The subpopulation of cells that survived serum-free cultiva-tion and paclitaxel treatment could highly express the cluster of differentiation 133/cluster of differentiation (CD133/CD326) mo-lecular markers and have features of stemness including differentiation ,high expression of cancer stem cells (CSCs)-associated genes and stronger capability of tumorigenesis .Conclusion The survived subpopulation that highly express the CD 133/CD326 molecu-lar markers presenting the characteristics of stemness in vitro and in vivo ,and could be used in future researches of biological functions .%目的:利用紫杉醇联合无血清培养完成对 A549肺腺癌始动细胞的富集并鉴定富集亚群的干细胞特性。方法对数生长期的 A549细胞经胰酶消化,干细胞培养基重悬,得到成球状生长的细胞;传至第2代时加入紫杉醇作用48 h ,离心去除死细胞和紫杉醇,换新鲜干细胞培养基培养,至存活细胞恢复克隆生长后鉴定其干细胞相关特性。结果紫杉醇联合无血清培养方式成功从 A549细胞中富集得到肿瘤干细胞,该群细胞高表达分化抗原簇蛋白133/人上皮细胞黏附分子(CD133/CD326),具有多向分化潜能、高表达干细胞相关基因及更强的致瘤能力,具备

  12. miRNA-200c对非小细胞肺癌A549细胞甲氨蝶呤耐药性的影响%Impact of miRNA-200c on Methotrexate Resistance of Non-small Cell Lung Cancer Cells A549

    单武林; 邓芳; 张晓蕾; 张婧; 韩丹丹; 万玲玲; 李明

    2016-01-01

    目的 探讨miRNA-200c(miR-200c)在非小细胞肺癌A549细胞耐甲氨蝶呤(MTX)(A549/MTX)中的影响及可能的作用机制.方法 通过实时荧光定量(qRT-PCR)法检测人肺癌亲本细胞NA549细胞、转染miR-200c模拟物(mimic)的A549/MTX细胞(A549/MTX-M)及转染miR-阴性对照(NC) A549/MTX细胞(A549/MTX-N)中miR-200c的表达水平.分别采用MTT法、锥虫兰染色及流式细胞术检测三组细胞对MTX的药物敏感度、细胞增殖能力及细胞凋亡变化,并采用qRT-PCR检测其P53和P21基因表达的变化.结果 miR-200c在A549细胞中的表达水平显著高于A549/MTX-N细胞;A549/MTX-M细胞miR-200c水平高于A549/MTX-N细胞;用不同浓度MTX刺激细胞,与A549/MTX-N细胞比较,A549/MTX-M细胞的增殖能力减弱、凋亡细胞增多,并呈剂量依赖性,差异均有统计学意义.转染miR-200c mimic后,P53和P21基因表达水平上升,与转染miR-NC细胞比较,差异有统计学意义.结论 miR-200c能够逆转A549/MTX细胞对MTX的耐药性,其作用机制可能是通过P53/P21信号转导通路诱导细胞凋亡来实现的.

  13. Anti-tumor activity of CrTX in human lung adenocarcinoma cell line A549

    Bin YE; Yan XIE; Zheng-hong QIN; Jun-chao WU; Rong HAN; Jing-kang HE

    2011-01-01

    Aim:To assess the cytotoxic effect of crotoxin (CrTX),a potent neurotoxin extracted from the venom of the pit viper Crotalus durissus terrificus,in human lung adenocarcinoma A549 cells and investigated the underlying mechanisms.Methods:A549 cells were treated with gradient concentrations of CrTX,and the cell cycle and apoptosis were analyzed using a flow cytometric assay.The changes of cellular effectors p53,caspase-3 and cleaved caspase-3,total P38MAPK and pP38MAPK were investigated using Western blot assays.A549 xenograft model was used to examine the inhibition of CrTX on tumor growth in vivo.Results:Treatment of A549 cells with CrTX (25-200 μg/mL) for 48 h significantly inhibited the cell growth in a dose-dependent manner (IC50=78 μg/mL).Treatment with CrTX (25 iJg/mL) for 24 h caused G1 arrest and induced cell apoptosis.CrTX (25 μg/mL) significantly increased the expression of wt p53,cleaved caspase-3 and phospho-P38MAPK.Pretreatment with the specific P38MAPK inhibitor SB203580 (5 μmol/L) significantly reduced CrTX-induced apoptosis and cleaved caspase-3 level,but G1 arrest remained unchanged and highly expressed p53 sustained.Intraperitoneal injection of CrTX (10 μg/kg,twice a week for 4 weeks) significantly inhibited A549 tumor xenograft growth,and decreased MVD and VEGF levels.Conclusion:CrTX produced significant anti-tumor effects by inducing cell apoptosis probably due to activation of P38MAPK and caspase-3,and by cell cycle arrest mediated by increased wt p53 expression.In addition,CrTX displayed anti-angiogenic effects in vivo.

  14. 小干扰RNA 抑制Pokemon表达对肺癌细胞A549和食管癌细胞EC109增殖的影响%Effects of Down-regulating Pokemon Expression by siRNA on Proliferation of Lung Cancer Cell A549 and Esophageal Cancer Cell EC109

    沈惠琳; 叶月芳; 丛德刚

    2014-01-01

    目的:探讨Pokemon特异性小干扰RNA对肺癌细胞A549和食管癌细胞EC109 增殖的影响.方法:瞬时转染Pokemon特异性小干扰RNA至肺癌细胞A549和食管癌细胞EC109,RT-PCR、Western Blot技术检测转染后细胞中Pokemon的mRNA和蛋白表达水平,检测细胞的增殖及细胞周期变化.结果:与空白组和阴性对照组相比,瞬时转染Pokemon小干扰RNA后,肺癌细胞A549和食管癌细胞EC109中Pokemon的mRNA水平均下降至25%~35%,蛋白水平亦明显下降.细胞增殖能力在培养24,48,72 h均显著降低(P<0.05).细胞周期分析显示转染Pokemon小干扰RNA后S期的比例显著高于siRNA阴性对照组(A549细胞:55.7%±2.5% vs 42.7%±0.6%,P<0.01;EC109细胞:67.7%±2.5% vs 52.0%±2.0%,P<0.01).G1期的比例显著低于siRNA阴性对照组(A549细胞:33.0%±2.0% vs 45.3%±1.5%,P<0.01;EC109细胞:30.7%±1.2% vs 44.0%±1.7%,P<0.01).两种细胞均阻滞于S期.结论:Pokemon小干扰RNA可抑制肺癌细胞A549和食管癌细胞EC109 的增殖.

  15. Wogonin has multiple anti-cancer effects by regulating c-Myc/SKP2/Fbw7α and HDAC1/HDAC2 pathways and inducing apoptosis in human lung adenocarcinoma cell line A549.

    Xin-mei Chen

    Full Text Available Wogonin is a plant monoflavonoid which has been reported to inhibit cell growth and/or induce apoptosis in various tumors. The present study examined the apoptosis-inducing activity and underlying mechanism of action of wogonin in A549 cells. The results showed that wogonin was a potent inhibitor of the viability of A549 cells. Apoptotic protein changes detected after exposure to wogonin included decreased XIAP and Mcl-1 expression, increased cleaved-PARP expression and increased release of AIF and cytochrome C. Western blot analysis showed that the activity of c-Myc/Skp2 and HDAC1/HDAC2 pathways, which play important roles in tumor progress, was decreased. Quantitative PCR identified increased levels of c-Myc mRNA and decreased levels of its protein. Protein levels of Fbw7α, GSK3β and Thr58-Myc, which are involved in c-Myc ubiquitin-dependent degradation, were also analyzed. After exposure to wogonin, Fbw7α and GSK3β expression decreased and Thr58-Myc expression increased. However, MG132 was unable to prevent c-Myc degradation. The present results suggest that wogonin has multiple anti-cancer effects associated with degradation of c-Myc, SKP2, HDAC1 and HDAC2. Its ability to induce apoptosis independently of Fbw7α suggests a possible use in drug-resistance cancer related to Fbw7 deficiency. Further studies are needed to determine which pathways are related to c-Myc and Fbw7α reversal and whether Thr58 phosphorylation of c-Myc is dependent on GSK3β.

  16. 甲氨蝶呤对映体诱导肺癌细胞耐药后引起血管内皮细胞分化差异的研究%Chiral selectivity in differentiation of lung cancer A549 cells to vascular endothelial cells after drug resistance induced by D-or L-methotrexate enantiomers

    何晓东; 胡世莲; 沈佐君; 陶绍能; 董林; 朱园园; 李明

    2009-01-01

    Objective To study the chiral selectivity in vascular endothelial differentiation in drug resistant lung cancer cells induced by high-dose L- or D-methotrexate (MTX) enantiomer. Methods Human lung cancer cells of the line A549 were co-cultured with high-dose (15 μmol/L) L- or D- MTX enantiomer so as to develop cancer cells resistant to MTX. MTT method was used to detect the drug resistant index. Flow cytometry was used to detect the expression of CD44, a transmembrane glycoprotein reflecting the migration ability of cells, CD31, a marker of vascular endothelium, and P-170 protein. Fifteen BALB/c nude mice were inoculated with the parent A549 cells, L-MTX-resistant A549 cells induced by L-MTX enantiomer, and D-MTX-resistant A549 cells induced by D-MTX enantiomer. Four weeks later the mice were killed to take out the tumor masses, Immunohistochemistry with CD34 staining was used to detect the microvascular density (MVD). Results The drug resistant index of the D-MTX induced drug resistant A549 cells was 20.1±2.3, significantly higher than that of the L-MTX-induced cells (12.4±1.2, P=0,000). The CD44 positive rate of the D-MTX induced A549 cells was 97.0%±0.9%, not significantly different from that of the L-MTX-induced A549 cells (96.7%±1.4%, P=0.544). The CD31 positive rate of the D-MTX induced A549 cells was 91.9%±3.2%, significantly higher than that of the L-MTX-induced A549 cells (32.9%±8.0%, P=0.000). The P-170 protein positive rate of the parent cells was 85.5%±4.6%, and the P-170 protein positive rate of the D-MTX-induced A549 cells was 87.0%±8.9%, significantly higher than that of the L-MTX-induced cells (71.5%±8.2%, P=0.002). The MVD of the D-MTX-indueed cells was 55.9±11.9, significantly higher than that of the L-MTX-induced cells (7. 2±1.7, P=0.000). MVD was significantly positively correlated with the CD31 level (r=0.462, P=0.007), and not correlated with P-170 protein and CD34 levels. Conclusion The MTX enantiomers have different chiral

  17. Differential expressions of VEGF and its receptor in methotrexate enantiomer-resistant lung cancer A549 cells%甲氨蝶呤对映体诱导的肺癌A549耐药细胞株中VEGF及其受体表达差异的研究

    董林; 何晓东; 陶绍能; 孙余婕; 李明; 沈佐君

    2009-01-01

    目的:通过探讨甲氨蝶呤(methotrexate, MTX)对映体耐药细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)及其受体表达的差异,以反映MTX对映体耐药细胞在肿瘤中血管形成能力的不同.方法:采用实时荧光定量PCR(real-time fluorogentic quantitative PCR,RFQ-PCR)技术分别检测D-(-)-MTX/A549组、L-(+)-MTX/A549组以及亲本细胞组中VEGF、血管内皮生长因子受体(vascular endothelial growth factor receptor,VEGFR) - 1和VEGFR - 2 mRNA的表达水平;皮下接种裸鼠,成瘤后取瘤体切片,行CD34免疫组织化学检测,新生微血管密度(microvessel density,MVD)计数;采用双层软琼脂克隆形成实验检测D-(-)-MTX/A549组、L-(+)-MTX/A549组细胞的克隆形成率.结果:RFQ-PCR检测结果显示,D-(-)-MTX/A549组VEGF、VEGFR-1和VEGFR-2 Ct值/β-actin Ct值分别为1.668±0.127、1.872±0.133和1.919±0.107,L-(+)-MTX/A549组的比值分别为2.035±0.185、2.221 ±0.157和2.255 ±0.140,亲本细胞组的比值为2.057±0.123、2.291±0.138和2.354±0.131;3种细胞植入裸鼠成瘤后切片免疫标记CD34统计MVD显示亲本细胞为3.29±1.11,L-(+)-MTX/A549细胞为8.00±2.14,D-(-)-MTX/A549细胞为57.88±13.87;软琼脂实验检测克隆形成率发现D-(-)-MTX/A549组为(0.625±0.088)%,L-(+)-MTX/A549组为(1.050±0.095)%,亲本细胞组为(1.250±0.248)%.研究结果显示D-(-)-MTX/A549组和L-(+)-MTX/A549组以及亲本细胞组之间差异均有统计学意义(P0.05).结论:D-(-)-MTX诱导肺腺癌A549耐药细胞在肿瘤血管形成上的能力大于L-(+)-MTX诱导的细胞.

  18. MicroRNA-200c depresses the migration and invasion of methotrexate-resistant lung cancer A549/MTX cells through EZH2/E-cad signaling pathway%微RNA-200c通过EZH2/E-cad信号通路抑制肺癌A549/MTX耐药细胞的迁移和侵袭能力

    张晓蕾; 邓芳; 单武林; 杨臣欢; 魏敏; 李程; 吴坤; 韩丹丹; 张婧

    2015-01-01

    目的:探讨微RNA-200c(microRNA-200c,miR-200c)对耐受甲氨蝶呤(methotrexate,MTX)的人非小细胞肺癌A549细胞迁移和侵袭能力的影响,及其可能的分子作用机制.方法:通过浓度递增结合低剂量持续诱导,获得耐受MTX的人肺癌A549/MTX细胞系后,观察诱导前后细胞的形态学改变.将miR-200c模拟物(mimic)转染A549/MTX细胞株后,分别采用细胞划痕愈合实验和Transwell细胞迁移和侵袭实验检测细胞的迁移和侵袭能力;再用实时荧光定量PCR法检测miR-200c过表达的A549/MTX细胞中人Zeste基因增强子同源物2 (human enhancer of Zeste homolog 2,EZH2)和E-钙黏蛋白(E-cadherin,E-cad)的mRNA表达水平.结果:肺癌A549/MTX耐药细胞构建成功.miR-200c minic转染后A549/MTX耐药细胞表达miR-200c水平比转染阴性片段组高6.41倍(P<0.05),表明转染成功.miR-200c mimic转染后A549/MTX细胞的迁移和侵袭能力显著降低(P值均< 0.05);而且A549/MTX细胞中EZH2 mRNA表达水平明显降低,而E-cad mRNA水平明显升高(P值均<0.05).结论:miR-200c高表达可以抑制A549/MTX耐药细胞的迁移和侵袭能力,其机制可能与其下调EZH2表达和上调E-cad水平有关.

  19. 人肺癌A549细胞系肿瘤干细胞的筛选和鉴定%Identiifcation and Isolation of Cancer Stem Cells from A549 Cells

    夏晖; 于长海; 张文; 张宝石; 赵英男; 方芳

    2013-01-01

    Background and objective Lung cancer stem cells are the root causes of lung cancer malignant phe-notype and potential therapeutic target, the aim of this study is to isolate and characterize the cancer stem cells in the lung adenoearcinomas cell line A549, so as to provide an experimental basis for further stem cell research. Methods hTe cancer stem cells were isolated from the lung adenoearcinomas cell line A549 using FACS. And the difference of colony formation, cell proliferation and tumorigenicity in vitro were also tested. hTe expression of CD133 and ABCG2 were evaluated by RT-PCR and Western blot. Results hTe percentage of SP cells was 5.93%of A549 and 0.32%of A549 atfer incubation with verapamil. hTe results showed that there were signiifcantly higher expression of CD133 and ABCG2 on SP cells than that of non-SP cells. And the ability of colony formation, cell proliferation and tumorigenicity in SP cell group were remarkably higher than that in non-SP cell group. Conclusion Our results suggested that the cancer stem cells with higher expression of CD133 and ABCG2 can be isolated from the lung adenoearcinomas cell line A549 using FACS and be used in the further research experiments.%背景与目的肺癌干细胞是肺癌恶性表型的根源和潜在的治疗靶点,从人肺癌A549细胞株中分离肺癌干细胞,观察特异性干细胞标志物分子的表达,为进一步的干细胞研究提供试验基础。方法接种肺癌A549细胞株,经流式细胞术,特异性筛选分离肺癌干细胞,观察克隆形成能力、细胞增殖能力和体外致瘤能力的差别,并分别用RT-PCR和Western blot的方法分析干细胞标志物分子CD133和ABCG2的表达。结果经过流式细胞仪成功分选了人肺腺癌A549细胞系SP细胞亚群,结果表明此SP细胞亚群约占A549细胞总数的5.93%,经维拉帕米处理后Hoechest33342阴性/弱阳性细胞含量下降为0.32%,SP细胞克隆形成能力,细胞增殖能力和

  20. Influence of Tamoxifen or the combination of Tamoxifen and Cisplatin on the growth of human lung adenocarcinoma A549 cells

    Yuxuan Che; Xiuhua Sun; Chaomei Huang; Jinbo Zhao 

    2014-01-01

    Objective:The experiment aims to investigate the influence of Tamoxifen and the combination of Tamoxifen and Cisplatin (DDP) on the growth of human lung adenocarcinoma A549 cel s. Methods:We treated human lung adenocarcinoma A549 cel s with dif erent concentrations of Tamoxifen, DDP and combination of DDP and Tamoxifen with non-toxicity for 72 h. Then we calculated the inhibition rate through MTT approach and detected the apoptosis rate by flow cytometry. The statistical analysis was performed with SPSS 13.0 software and statistical dif erences were determined by one-way ANOVA. The data were expressed as the mean ± standard deviation and al experiments were performed in three times. The value of P0.05). 2. As the increase concentration of Tamoxifen, the S stage and G2/M of the A549 cel s decreased while the G0/G1 increased. The apoptosis rate of Tamoxifen with 0 µmol/L, 0.1 µmol/L, 1 µmol/L and 10 µmol/L on the A549 cel s were 6.51%, 8.91%, 17.97%and 42.7%, respectively. 3. The inhibition rates of combination of Tamoxifen with 1 µmol/L and DDP with 1.25 µg/mL, 2.5 µg/mL, 5 µg/mL, 10 µg/mL and 20 µg/mL on the A549 cel s were 40.4%, 54.4%, 72.9%, 86.1%and 92.4%, respectively (P<0.05). Conclusion:Tamoxifen can inhibit the proliferation of human lung adenocarcinoma A549 cel s and induce the apoptosis of the A549 cel s. The combination of Tamoxifen with non-toxicity and DDP can improve the sensitivity of chemotherapy on the A549 cel s.

  1. The Anti-Lung Cancer Activities of Steroidal Saponins of P. polyphylla Smith var. chinensis (Franch. Hara through Enhanced Immunostimulation in Experimental Lewis Tumor-Bearing C57BL/6 Mice and Induction of Apoptosis in the A549 Cell Line

    Rui-Ping Wang

    2013-10-01

    Full Text Available P. polyphylla Smith var. chinensis (Franch. Hara (PPSCFH has been used as medicinal Paris for the prevention and treatment of cancers in China for thousands of years. Its main components, steroidal saponins (PRS, have been confirmed to inhibit tumor growth. In the present study, the immunostimulation of PRS was investigated in Lewis bearing-C57BL/6 mice while the induction of apoptosis in A549 cells was also studied. The treatment with PRS (2.5, 5.0 and 7.5 mg/kg significantly inhibited tumor, volume, and weight in the C57BL/6 mice. The rates of inhibition of PRS (at 2.5, 5.0 and 7.5 mg/kg were 26.49 ± 17.30%, 40.32 ± 18.91% and 54.94 ± 16.48%, respectively. The spleen and thymus indexes were increased remarkably, while the levels of inflammatory cytokines including TNF-α, IL-8 and IL-10 in serum were decreased according to ELISA assays. For A549 cells, Hoechst 33342 staining and annexin V/PI by flow cytometry showed that PRS (0.25, 0.50 and 0.75 mg/mL induced nuclear changes of A549 cells with DNA condensation and fragmentations of chromatin, as well as inducing apoptosis. Furthermore, PRS could also attenuate the over-generation of intracellular ROS. Western blotting analysis showed a significant decrease on the expressions of proinflammatory cytokines MCP-1, IL-6 and TGF-β1, as well as cell adhesion molecule ICAM-1, by treatment with PRS. Our results demonstrated that the inhibition of PRS on tumor growth might be associated with the amelioration of inflammation responses, induction of apoptosis, as well as the decrease of ROS. These results suggested that PRS implied a potential therapeutic effect in the lung cancer treatment.

  2. 叶酰聚谷氨酸合成酶基因在甲氨蝶呤对映体获得性耐药A549细胞株中的表达差异%Differential gene expression of folylpolyglutamate synthetase in cytoplasm and mitochondria in acquired methotrexate enantiomers resistant to lung cancer A549 cell lines

    周红艳; 何晓东; 孙余婕; 凡任芝; 孙利; 沈佐君

    2011-01-01

    目的 研究不同甲氨蝶呤(MTX)对映体耐药与叶酰聚谷氨酸合成酶(FPGS)基因水平表达的关系.方法 用大剂量冲击递增结合低剂量持续诱导法诱导获得两组含不同构型15~55μmol/L浓度的MTX对映体[L-(+)-MTX和D-(-)-MTX]耐药的细胞系,细胞为人源非小细胞性肺癌A549细胞,用四甲基偶氮唑盐(MTT)法检测各细胞系的耐药指数;用实时荧光定量聚合酶链反应(RFQ-PCR)方法检测这两组各细胞系中胞质型FPGS(cFPGS)和线粒体型FPGS(mFPGS)基因的相对含量.结果 D-(-)-MTX耐药细胞组耐药指数高于L-(+)-MTX耐药细胞组(32.7±9.3比11.5±2.9,P<0.05),L-(+)-MTX/A549细胞系耐药指数均在5~15之间,为中度耐药,而D-(-)-MTX/A549细胞系耐药指数均>15,为高度耐药.在D-(-)-MTX和L-(+)-MTX两组耐药细胞系中,mFPGS表达水平仪在MTX为15 μmol/L时差异无统计学意义,在MTX其他各浓度点两组间差异均有统计学意义(25 μmol/L:2.3±0.9比1.3±0.7,35 μnol/L:2.6±0.3比1.1±0.9,45 μmol/L:1.4±0.8比1.0±1.0,55 μmol/L;1.0±0.2比0.2±0.1均P<0.05);cFPGS表达水平在MTX为15μmol/L时两组间差异也同样无统计学意义,在25~55 μmol/L浓度区间内D-(-)-MTX/A549细胞系的cFPGS表达与耐药指数呈现高度负相关(r=-0.95,P<0.05).结论 在A549细胞中MTX对映体初次剂量15 μmol/L冲击法诱导获得的对映体耐药与再次接受更大剂量(≥25 μmol/L)MTX诱导获得耐药的机制不同,D-(-)-MTX/A549耐药细胞系表现为更高的耐药性,提示临床使用MTX时应考虑该药物存在手性对映体问题.%Objective To investigate the relationship between the resistance of methotrexate (MTX) enantiomer and the gene expression levels of folylpolyglutamate synthetase (FPGS).Methods The cell lines of MTX enantiomer resistance from 15 -55 μmol/L were obtained when the A549 cell lines were exposed intermittently and progressively to an incremental dose of each MTX enantiomer.The resistant

  3. 利用PCR-SSP法研究肺腺癌细胞系A549、Calu-6的HLA-ABDR等位基因%Study on HLA-ABDR alleles in A549 and Calu-6 lung cancer cell lines with PCR-SSP

    邓波; 林一丹; 王如文; 蒋耀光

    2006-01-01

    背景和目的已有的研究表明人类白细胞抗原(HLA)在抗原呈递及T细胞识别抗原的过程中起关键作用,此外还与肿瘤细胞的免疫杀伤及免疫逃避有着密切的关系.本研究探讨了人肺腺癌细胞系A549、Calu-6中HLA-A、HLA-B、HLA-DR等位基因的存在状况.方法分离A549、Calu-6细胞DNA,分别行PCR-SSP法扩增、电泳后紫外透射扫描,根据反应格局表对HLA-A、HLA-B、HLA-DR进行判定.结果A549与Calu-6细胞中HLA-A、HLA-B基因较杂合子均有缺失,而HLA-DR基因无缺失.A549细胞HLA-ABDR的基因分型为HLA-A30、HLA-B44、HLA-DR7/HLA-DR53.Calu-6细胞HLA-ABDR的基因分型为HLA-A01、HLA-B08、HLA-DR17/HLA-DR52.结论肺腺癌中存在HLA-Ⅰ和HLA-Ⅱ基因.HLA-Ⅰ基因可能在肿瘤细胞传代过程中发生选择性丢失,而HLA-DR基因完整保留.检测肿瘤HLA对了解其免疫学行为及建立肿瘤特异性杀伤淋巴细胞(CTL)模型具有重要意义.

  4. DNA damage response signaling in lung adenocarcinoma A549 cells following gamma and carbon beam irradiation

    Ghosh, Somnath [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India); Narang, Himanshi, E-mail: himinarang@gmail.com [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India); Sarma, Asitikantha [Radiation Biology Laboratory, Inter University Accelerator Centre, Aruna Asaf Ali Marg, New Delhi 110 067 (India); Krishna, Malini [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India)

    2011-11-01

    Carbon beams (5.16 MeV/u, LET = 290 keV/{mu}m) are high linear energy transfer (LET) radiation characterized by higher relative biological effectiveness than low LET radiation. The aim of the current study was to determine the signaling differences between {gamma}-rays and carbon ion-irradiation. A549 cells were irradiated with 1 Gy carbon or {gamma}-rays. Carbon beam was found to be three times more cytotoxic than {gamma}-rays despite the fact that the numbers of {gamma}-H2AX foci were same. Percentage of cells showing ATM/ATR foci were more with {gamma}-rays however number of foci per cell were more in case of carbon irradiation. Large BRCA1 foci were found in all carbon irradiated cells unlike {gamma}-rays irradiated cells and prosurvival ERK pathway was activated after {gamma}-rays irradiation but not carbon. The noteworthy finding of this study is the early phase apoptosis induction by carbon ions. In the present study in A549 lung adenocarcinoma, authors conclude that despite activation of same repair molecules such as ATM and BRCA1, differences in low and high LET damage responses might be due to their distinct macromolecular complexes rather than their individual activation and the activation of cytoplasmic pathways such as ERK, whether it applies to all the cell lines need to be further explored.

  5. Construction of Eukaryotic Expression Vector of Human CC10 Gene and Expression of CC10 Protein in Lung Adenocarcinoma A549 Cell Line

    2005-01-01

    A mammalian expression plasmid pcDNA3.1-hCC10 was constructed and identified, then CC10 protein expression in A549 lung cancer cell line was detected. A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by using RT-PCR and cloned into expression plasmid cDNA3.1, and the recombinant plasmid was identified by employing double digestion restriction enzymes HindⅢ and BamH Ⅰ and the cDNA sequence was assayed by the Sanger dideoxymediated chain termination method. The segment was then transfected into the A549 lung cancer cell line. The protein expression of CC10 was detected by immunofluorescence and Western blot.Our results showed that the cDNA fragment included the entire coding region (273 bp). The recombinant eukaryotic cell expression vector of pcDNA3.1-hCC10 was successfully constructed, and the sequence of the insert was identical to the published sequence. A549 cells line transfected with the pcDNA3.1-hCC10 expressed high level of CC10 protein. The recombinant plasmid cDNA3. 1hCC10 may serve as an effective tool for the study of tumorogenesis and tumor treatment.

  6. Silica nanoparticles and biological dispersants: genotoxic effects on A549 lung epithelial cells

    Silica nanoparticle exposure could be intentional (e.g. medical application or food) or accidental (e.g. occupational inhalation). On entering the body, particles become coated with specific proteins depending on the route of entry. The ability of silica particles of different size and charge (non-functionalized 50 and 200 nm and aminated 50 and 200 nm) to cause genotoxic effects in A549 lung epithelial cells was investigated. Using the modified comet assay and the micronucleus assay, we examined the effect of suspending the particles in different dispersion media [RPMI or Hanks’ balanced salt solution (HBSS), supplemented with bovine serum albumin (BSA), lung lining fluid (LLF) or serum] to determine if this influenced the particle’s activity. Particle characterisation suggested that the particles were reasonably well dispersed in the different media, with the exception of aminated 50 nm particles which showed evidence of agglomeration. Plain 50, 200 nm and aminated 50 nm particles caused significant genotoxic effects in the presence of formamidopyrimidine-DNA glycosylase when dispersed in HBSS or LLF. These effects were reduced when the particles were dispersed in BSA and serum. There was no significant micronucleus formation produced by any of the particles when suspended in any of the dispersants. The data suggest that silica particles can produce a significant genotoxic effect according to the comet assay in A549 cells, possibly driven by an oxidative stress-dependent mechanism which may be modified depending on the choice of dispersant employed

  7. Silica nanoparticles and biological dispersants: genotoxic effects on A549 lung epithelial cells

    Brown, David M., E-mail: d.brown@hw.ac.uk [Heriot-Watt University, Nanosafety Research Group, School of Life Sciences (United Kingdom); Varet, Julia, E-mail: julia.varet@IOM-world.org [Institute of Occupational Medicine (United Kingdom); Johnston, Helinor, E-mail: h.johnston@hw.ac.uk; Chrystie, Alison; Stone, Vicki, E-mail: v.stone@hw.ac.uk [Heriot-Watt University, Nanosafety Research Group, School of Life Sciences (United Kingdom)

    2015-10-15

    Silica nanoparticle exposure could be intentional (e.g. medical application or food) or accidental (e.g. occupational inhalation). On entering the body, particles become coated with specific proteins depending on the route of entry. The ability of silica particles of different size and charge (non-functionalized 50 and 200 nm and aminated 50 and 200 nm) to cause genotoxic effects in A549 lung epithelial cells was investigated. Using the modified comet assay and the micronucleus assay, we examined the effect of suspending the particles in different dispersion media [RPMI or Hanks’ balanced salt solution (HBSS), supplemented with bovine serum albumin (BSA), lung lining fluid (LLF) or serum] to determine if this influenced the particle’s activity. Particle characterisation suggested that the particles were reasonably well dispersed in the different media, with the exception of aminated 50 nm particles which showed evidence of agglomeration. Plain 50, 200 nm and aminated 50 nm particles caused significant genotoxic effects in the presence of formamidopyrimidine-DNA glycosylase when dispersed in HBSS or LLF. These effects were reduced when the particles were dispersed in BSA and serum. There was no significant micronucleus formation produced by any of the particles when suspended in any of the dispersants. The data suggest that silica particles can produce a significant genotoxic effect according to the comet assay in A549 cells, possibly driven by an oxidative stress-dependent mechanism which may be modified depending on the choice of dispersant employed.

  8. canstatin基因转染对肺癌A549细胞和血管内皮细胞增殖与凋亡的影响%Effects of canstatin gene transfection on growth and apoptosis of lung cancer A549 cells and HUV-ECC cells

    陆卫忠; 黄桂君; 钱桂生; 李玉英; 余时沧; 李瑾

    2005-01-01

    背景与目的肿瘤的生长和转移需要大量新血管的生成,人血管能抑素(canstatin)是新近发现的高效内源性血管生成抑制剂,其抑制血管内皮细胞的作用已引起人们广泛关注.本研究的目的是探讨canstatin基因在人肺癌A549细胞和人脐静脉内皮细胞HUV-ECC中的表达及意义.方法将canstatin基因通过电穿孔的方法转染人肺癌细胞A549和人脐静脉内皮细胞HUV-ECC,行G418筛选获得转基因细胞克隆.用SDS-PAGE电泳检测canstatin蛋白在转基因细胞培养上清液中的表达,以流式细胞仪分析细胞周期,并比较转基因和未转基因细胞的生长特性.结果 canstatin在转染组A549细胞和ECC细胞表达并分泌至上清液中.canstatin基因转染组ECC的凋亡率(16.04%)显著高于空载体组(0.43%)和亲代细胞组(2.92%)(P0.05),细胞生长也未受明显影响.结论 canstatin能特异地抑制内皮细胞增殖,并诱导内皮细胞凋亡.

  9. 肺癌细胞A549抗原相关旋毛虫Tsp06172基因的克隆及原核表达%Cloning and prokaryotic expression of the Tsp06172 gene of Trichinella spiralis in A549 lung cancer cells

    高江明; 徐晓芳; 吕萌; 左绍志; 宫鹏涛; 杨举; 李赫; 李建华; 张国才

    2013-01-01

    目的 克隆旋毛虫(Trichinella spiralis)与肺癌细胞A549相关抗原Tsp06172基因,并进行原核表达. 方法 采用RT-PCR方法扩增Tsp06172基因,连接原核表达载体pET-28a,转化入感受态细胞BL21,IPTG诱导表达,经SDS-PAGE和Western blot鉴定表达产物. 结果 重组表达质粒经双酶切及测序鉴定正确.表达分子质量单位约为16 ku的融合蛋白.Western blot检测融合蛋白能被抗A549细胞的多克隆抗体识别. 结论 构建的原核表达载体pET-28a Tsp06172表达具有A549细胞反应原性的蛋白,为旋毛虫Tsp06172重组蛋白功能的研究了奠定基础.%Objective To clone and express the Trichinella spiralis Tsp06172 gene in BL21. Methods The Tsp06172 gene was amplified with RT-PCR and then subcloned into the prokaryotic expression vector pET-28a. BL21 containing the recombinant plasmid pET-28a-Tsp06172 was induced with IPTG. The fusion protein was detected and i-dentified with SDS-PAGE and Western blotting. Results The recombinant expression plasmid was successfully constructed. After induction in an E. coli system. SDS-PAGE results showed that a fusion protein of about 16 ku was successfully expressed. Western blotting indicated that the fusion protein was readily recognized by polyclonal antibodies from A549 cells. Conclusion The recombinant expression plasmid pET-28a-Tsp06172 expressed the corresponding protein in BL21. This finding lays the foundation for research into the function of the Tsp06172 protein.

  10. Direct electric current treatment modifies mitochondrial function and lipid body content in the A549 cancer cell line.

    Holandino, Carla; Teixeira, Cesar Augusto Antunes; de Oliveira, Felipe Alves Gomes; Barbosa, Gleyce Moreno; Siqueira, Camila Monteiro; Messeder, Douglas Jardim; de Aguiar, Fernanda Silva; da Veiga, Venicio Feo; Girard-Dias, Wendell; Miranda, Kildare; Galina, Antonio; Capella, Marcia Alves Marques; Morales, Marcelo Marcos

    2016-10-01

    Electrochemical therapy (EChT) entails treatment of solid tumors with direct electric current (DC). This work evaluated the specific effects of anodic flow generated by DC on biochemical and metabolic features of the A549 human lung cancer cell line. Apoptosis was evaluated on the basis of caspase-3 activity and mitochondrial transmembrane potential dissipation. Cell morphology was analyzed using transmission electron microscopy, and lipid droplets were studied through morphometric analysis and X-ray qualitative elemental microanalysis. High-resolution respirometry was used to assess mitochondrial respiratory parameters. Results indicated A549 viability decreased in a dose-dependent manner with a prominent drop between 18 and 24h after treatment (pbodies' number, quantified by morphometrical analyses. Conversely, 24h after treatment, the cells presented a two-fold increase of residual oxygen consumption, accounting for 45.3% of basal oxygen consumption. These results show remarkable alterations promoted by anodic flow on human lung cancer cells which are possibly involved with the antitumoral effects of EChT. PMID:27243447

  11. Screening radiosensitizing-related genes mediated by elemene in lung adenocarcinoma A549 cells by using gene chip

    Objective: To screen radiosensitizing-related genes mediated by elemene in lung adenocarcinoma A549 cells by using gene chip. Methods: MTT test was used to calculate the IC50 of elemene. (1) The effect of radiosensitivity was detected by colony forming assay. A549 cells were divided into 2 groups: radiation group and radiation + elemene group. Oligonucleotide chip was used to screen the gene expression changes of A549 cells from these 2 groups. The up-regulated gene Egr-1 and the down-regulated gene CyclinD1 were selected to undergo RT-PCR so as to confirm the reliability of the result. Results: MTT test showed the elemene inhibited the proliferation of the A549 cells dose-dependently. The IC50 value of elemene on the A549 cells was 120 mg/L. (2) 10 mg/L elemene had radiosensitising effect on A549 cells.The values of SERD0 and SERDq obtained from the survival curve were (1.54±0. 20) and (1.43±0.15) respectively. Gene chip screened 122 differentially-expressed genes, including 89 up-regulated genes and 33 down-regulated genes. (3) These altered genes could be related to cell structure, substance metabolism,cell proliferation, cell differentiation, signal transduction, material transport, DNA repair, apoptosis, immune response and so forth. The RT-PCR results of Egr-1 and Cyclin D1 were consistent with the gene chip analysis. Conclusions: The mechanism of elemene enhancing the radiosensitivity of lung adenocarcinoma A549 cells is the result of participation and collaboration of multiple genes. Further study of the newly-discovered differentially-expressed gene helps find out new radiosensitizational targets of elemene. (authors)

  12. Lung Cancer

    ... spreads in different ways, and each is treated differently. Non-small cell lung cancer is more common than small cell lung cancer. Small cell lung cancer grows more quickly and is more likely to spread to other organs in the body. Learn more about non-small cell lung cancer. Learn ...

  13. Platinum(II) phenanthroimidazole G-quadruplex ligand induces selective telomere shortening in A549 cancer cells.

    Mancini, Johanna; Rousseau, Philippe; Castor, Katherine J; Sleiman, Hanadi F; Autexier, Chantal

    2016-02-01

    Telomere maintenance, achieved by the binding of protective shelterin capping proteins to telomeres and by either telomerase or a recombination-based alternative lengthening of telomere (ALT) mechanism, is critical for cell proliferation and survival. Extensive telomere shortening or loss of telomere integrity activates DNA damage checkpoints, leading to cell senescence or death. Although telomerase upregulation is an attractive target for anti-cancer therapy, the lag associated with telomere shortening and the potential activation of ALT pose a challenge. An alternative approach is to modify telomere interactions with binding proteins (telomere uncapping). G-quadruplex ligands stabilize structures generated from single-stranded G-rich 3'-telomere end (G-quadruplex) folding, which in principle, cannot be elongated by telomerase, thus leading to telomere shortening. Ligands can also mediate rapid anti-proliferative effects by telomere uncapping. We previously reported that the G-quadruplex ligand, phenylphenanthroimidazole ethylenediamine platinum(II) (PIP), inhibits telomerase activity in vitro[47]. In the current study, a long-term seeding assay showed that PIP significantly inhibited the seeding capacity of A549 lung cancer cells and to a lesser extent primary MRC5 fibroblast cells. Importantly, treatment with PIP caused a significant dose- and time-dependent decrease in average telomere length of A549 but not MRC5 cells. Moreover, cell cycle analysis revealed a significant increase in G1 arrest upon treatment of A549 cells, but not MRC5 cells. Both apoptosis and cellular senescence may contribute to the anti-proliferative effects of PIP. Our studies validate the development of novel and specific therapeutic ligands targeting telomeric G-quadruplex structures in cancer cells. PMID:26724375

  14. Modification of radio- and thermo-sensitivity by amrubicin or amrubicinol in human lung adenocarcinoma A549 cells

    Amrubicin (AMR) is a totally synthetic 9-aminoanthracyclin anticancer drug. It is considered that AMR is an inhibitor of DNA topoisomerase II as the case of another anthracyclin anticancer drug, adriamycin (ADM) (1), which has significant antitumor activity against a broad spectra of human malignancies. The antitumor activity of AMR was found superior to that of ADM in experimental therapeutic models of human tumor xenografts (nude mouse). AMR was converted in vivo to major metabolite, amrubicinol (AMROH), which was markedly more effective cytotoxic agent than the mother compound. In the clinical studies currently conducted on malignant lymphoma, non-small or small cell lung carcinoma, the activity of AMR was shown very promising. However, the interactive cytocidal effects of the combined treatment with AMR or AMROH and radiation or hyperthermia are under investigation. In the present study, we examined chemical modification of radio- and thermo-sensitivity by AMR or AMROH in cultured human lung adenocarcinoma A549 cells. Sublethal damage repair (SLDR) was inhibited by the pretreatment with AMR or AMROH followed by X-irradiation. This finding suggests the possibility of the combined treatment of AMR or AMROH and X-irradiation as clinical cancer therapy strategy, since the doses in the routine clinical radiotherapy is ranged with a sublethal dose of 2 Gy. We also found that SLDR was inhibited by the pretreatment with AMR or AMROH followed by hyperthermia. We will discuss about clinical adoption of the combined treatment with AMR or AMROH and radiation or hyperthermia

  15. 莪术油对人肺腺癌细胞A549增殖的影响%Effect of Zedoary Turmeric Oil on Proliferation in Human Lung Adenocarcinoma Cell Line A549

    王晓波; 牛建昭; 崔巍; 刘飒; 杨长福; 赵丕文; 唐炳华

    2011-01-01

    目的 探讨莪术油对人肺腺癌细胞A549增殖的抑制作用.方法 体外培养肺腺癌细胞A549,MTT比色法测定莪术油对A549细胞作用24、48、72 h后抑制率;流式细胞术分析莪术油对A549细胞作用24 h后细胞周期的变化;Annexin V-FITC/PI双染检测莪术油对A549细胞作用24 h后细胞凋亡与坏死情况.结果 莪术油对A549细胞增殖的抑制率随时间延长明显升高,随药物浓度增加抑制作用增强;莪术油对A549细胞作用24 h后,细胞周期停滞在G0/G1期,阻止其进入S期;细胞的早期凋亡、晚期凋亡和坏死比例随着莪术油浓度的增加而增加,且坏死细胞的比例高于凋亡细胞.结论 莪术油对A549细胞的增殖具有抑制作用,并呈时间、浓度依赖,其作用是通过阻滞细胞周期及诱导凋亡和坏死采实现的.%Objective To explore the inhibiting effect of Zedoary turmeric oil on the proliferation of A549 cell line. Methods Lung adenocarcinoma cell line A549 was cultured in vitro. The inhibition rate of Zedoary turmeric oil on the proliferation of lung adenocarcinoma cell line A549 for 24, 48, 72 h were determined by MTT colorimetric assay. The cell cycle of lung adenocarcinoma cell line A549 stimulated by Zedoary turmeric oil for 24 h was analyzed by flow cytometry. The apoptosis and necrosis of lung adenocarcinoma cell line A549 stimulated by Zedoary turmeric oil for 24 h was tested by Annexin V-FITC/PI assay. Results MTT assay indicated that the inhibition rate of Zedoary turmeric oil on the proliferation of lung adenocarcinoma cell line A549 increased significantly with the growing of time and concentration. Further analysis by flow cytometry indicated that Zedoary turmeric oil stimulating the A549 cells for 24 h led to Go/Gi phase arrest and blocked S phase entry. Meanwhile cells in early apoptosis, late apoptosis and necrosis were increased, and the percentage of necrotic cells was more than apoptotic cells with the increase of

  16. Tumor-targeting magnetic lipoplex delivery of short hairpin RNA suppresses IGF-1R overexpression of lung adenocarcinoma A549 cells in vitro and in vivo

    Wang, Chunmao; Ding, Chao; Kong, Minjian [Department of Cardiothoracic Surgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009 (China); Dong, Aiqiang, E-mail: dr_dongaiqiang@sina.com [Department of Cardiothoracic Surgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009 (China); Qian, Jianfang; Jiang, Daming; Shen, Zhonghua [Department of Cardiothoracic Surgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009 (China)

    2011-07-08

    Highlights: {yields} We compared lipofection with magnetofection about difference of transfection efficiency on delivery a therapeutic gene in vitro and in vivo. {yields} We investigated the difference of shRNA induced by magnetofection and lipofection into A549 cell and subcutaneous tumor to knockdown IGF-1R overexpressed in A549 cell and A549 tumor. {yields} We investigated in vivo shRNA silenced IGF-1R overexpression 24, 48, and 72 h after shRNA intravenous injection into tumor-bearing mice by way of magnetofection and lipofection. {yields} Our results showed that magnetofection could achieve therapeutic gene targeted delivery into special site, which contributed to targeted gene therapy of lung cancers. -- Abstract: Liposomal magnetofection potentiates gene transfection by applying a magnetic field to concentrate magnetic lipoplexes onto target cells. Magnetic lipoplexes are self-assembling ternary complexes of cationic lipids with plasmid DNA associated with superparamagnetic iron oxide nanoparticles (SPIONs). Type1insulin-like growth factor receptor (IGF-1R), an important oncogene, is frequently overexpressed in lung cancer and mediates cancer cell proliferation and tumor growth. In this study, we evaluated the transfection efficiency (percentage of transfected cells) and therapeutic potential (potency of IGF-1R knockdown) of liposomal magnetofection of plasmids expressing GFP and shRNAs targeting IGF-1R (pGFPshIGF-1Rs) in A549 cells and in tumor-bearing mice as compared to lipofection using Lipofectamine 2000. Liposomal magnetofection provided a threefold improvement in transgene expression over lipofection and transfected up to 64.1% of A549 cells in vitro. In vitro, IGF-1R specific-shRNA transfected by lipofection inhibited IGF-1R protein by 56.1 {+-} 6% and by liposomal magnetofection by 85.1 {+-} 3%. In vivo delivery efficiency of the pGFPshIGF-1R plasmid into the tumor was significantly higher in the liposomal magnetofection group than in the

  17. Tumor-targeting magnetic lipoplex delivery of short hairpin RNA suppresses IGF-1R overexpression of lung adenocarcinoma A549 cells in vitro and in vivo

    Highlights: → We compared lipofection with magnetofection about difference of transfection efficiency on delivery a therapeutic gene in vitro and in vivo. → We investigated the difference of shRNA induced by magnetofection and lipofection into A549 cell and subcutaneous tumor to knockdown IGF-1R overexpressed in A549 cell and A549 tumor. → We investigated in vivo shRNA silenced IGF-1R overexpression 24, 48, and 72 h after shRNA intravenous injection into tumor-bearing mice by way of magnetofection and lipofection. → Our results showed that magnetofection could achieve therapeutic gene targeted delivery into special site, which contributed to targeted gene therapy of lung cancers. -- Abstract: Liposomal magnetofection potentiates gene transfection by applying a magnetic field to concentrate magnetic lipoplexes onto target cells. Magnetic lipoplexes are self-assembling ternary complexes of cationic lipids with plasmid DNA associated with superparamagnetic iron oxide nanoparticles (SPIONs). Type1insulin-like growth factor receptor (IGF-1R), an important oncogene, is frequently overexpressed in lung cancer and mediates cancer cell proliferation and tumor growth. In this study, we evaluated the transfection efficiency (percentage of transfected cells) and therapeutic potential (potency of IGF-1R knockdown) of liposomal magnetofection of plasmids expressing GFP and shRNAs targeting IGF-1R (pGFPshIGF-1Rs) in A549 cells and in tumor-bearing mice as compared to lipofection using Lipofectamine 2000. Liposomal magnetofection provided a threefold improvement in transgene expression over lipofection and transfected up to 64.1% of A549 cells in vitro. In vitro, IGF-1R specific-shRNA transfected by lipofection inhibited IGF-1R protein by 56.1 ± 6% and by liposomal magnetofection by 85.1 ± 3%. In vivo delivery efficiency of the pGFPshIGF-1R plasmid into the tumor was significantly higher in the liposomal magnetofection group than in the lipofection group. In vivo IGF-1R

  18. Effects of X-ray irradiation on expression of Pokemon gene in A549 cells of human lung adenocarcinoma

    Objective: To study the dose-and time-effects of X-ray irradiation on the expression of Pokemon gene in A549 cells of human lung adenocarcinoma. Methods: A549 cells were cultured in vitro and exposed to X-rays with the doses of 2, 4, 6 and 8 Gy, respectively. Untreated A549 cells were used as control group. The relative levels of Pokemon mRNA expression in the cells were detected by using quantitative real-time PCR at 2, 4, 8, 12, 24, 48 and 72 h after irradiation. Results: The Pokemon mRNA expression levels decreased in the early period after irradiation (except 2 and 4 h after irradiation in 2 Gy group) and then increased in the later stage (48 h after irradiation) with significant statistical differences at the most time points in comparison with the control group (t=3.40-154.76, P<0.05). Conclusions: Higher doses of X-rays may degrade the expression of Pokemon mRNA in the human A549 cells and induce apoptosis in the early period, hut also may upgrade its expression in the later period, which might be correlated with the cell cycle regulation and DNA damage repair in the A549 cells. (authors)

  19. Lung Cancer

    Lung cancer is one of the most common cancers in the world. It is a leading cause of ... in the United States. Cigarette smoking causes most lung cancers. The more cigarettes you smoke per day and ...

  20. Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells

    Torshabi, Maryam; Faramarzi, Mohammad Ali; Tabatabaei Yazdi, Mojtaba; Ostad, Seyyed Naser; Gharemani, Mohammad Hosein

    2011-01-01

    Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS an...

  1. The effect of sulforaphane on the cell cycle, apoptosis and expression of cyclin D1 and p21 in the A549 non-small cell lung cancer cell line.

    Żuryń, Agnieszka; Litwiniec, Anna; Safiejko-Mroczka, Barbara; Klimaszewska-Wiśniewska, Anna; Gagat, Maciej; Krajewski, Adrian; Gackowska, Lidia; Grzanka, Dariusz

    2016-06-01

    Sulforaphane (SFN) is present in plants belonging to Cruciferae family and was first isolated from broccoli sprouts. Chemotherapeutic and anticarcinogenic properties of sulforaphane were demonstrated, however, the underlying mechanisms are not fully understood. In this study we evaluated the expression of cyclin D1 and p21 protein in SFN-treated A549 cells and correlated these results with the extent of cell death and/or cell cycle alterations, as well as determined a potential contribution of cyclin D1 to cell death. A549 cells were treated with increasing concentrations of SFN (30, 60 and 90 µM) for 24 h. Morphological and ultrastructural changes were observed using light, transmission electron microscope and videomicroscopy. Image-based cytometry was applied to evaluate the effect of SFN on apoptosis and the cell cycle. Cyclin D1 and p21 expression was determined by flow cytometry, RT-qPCR and immunofluorescence. siRNA was used to evaluate the role of cyclin D1 in the process of suforaphane-induced cell death. We found that the percentage of cyclin D1-positive cells decreased after the treatment with SFN, but at the same time mean fluorescence intensity reflecting cyclin D1 content was increased at 30 µM SFN and decreased at 60 and 90 µM SFN. Percentage of p21-positive cells increased following the treatment, with the highest increase at 60 µM SFN, at which concentration mean fluorescence intensity of this protein was also significantly increased. The 30-µM dose of SFN induced an increased G2/M phase population along with a decreased polyploid fraction of cells, which implies a functional G2/M arrest. The major mode of cell death induced by SFN was necrosis and, to a lower degree apoptosis. Transfection with cyclin D1-siRNA resulted in significantly compromised fraction of apoptotic and necrotic cells, which suggests that cyclin D1 is an important determinant of the therapeutic efficiency of SFN in the A549 cells. PMID:27035641

  2. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells

    Wu Xinjiang [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Kassie, Fekadu [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Mersch-Sundermann, Volker [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany)]. E-mail: Volker.mersch-sundermann@uniklinikum-giessen.de

    2005-11-11

    Diallyl disulfide (DADS), an oil soluble constituent of garlic (Allium sativum), has been reported to cause antimutagentic and anticarcinogenic effects in vitro and in vivo by modulating phases I and II enzyme activities. In recent years, several studies suggested that the chemopreventive effects of DADS can also be attributed to induction of cell cycle arrest and apoptosis in cancer cells. In the present study, we reported that DADS-induced cell cycle arrest at G2/M and apoptosis in human A549 lung cancer cells in a time- and dose-dependent manner. Additionally, a significant increase of intracellular reactive oxygen species (ROS) was induced in A549 cells less than 0.5 h after DADS treatment, indicating that ROS may be an early event in DADS-modulated apoptosis. Treatment of A549 cells with N-acetyl cysteine (NAC) completely abrogated DADS-induced cell cycle arrest and apoptosis. The result indicated that oxidative stress modulates cell proliferation and cell death induced by DADS.

  3. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells

    Diallyl disulfide (DADS), an oil soluble constituent of garlic (Allium sativum), has been reported to cause antimutagentic and anticarcinogenic effects in vitro and in vivo by modulating phases I and II enzyme activities. In recent years, several studies suggested that the chemopreventive effects of DADS can also be attributed to induction of cell cycle arrest and apoptosis in cancer cells. In the present study, we reported that DADS-induced cell cycle arrest at G2/M and apoptosis in human A549 lung cancer cells in a time- and dose-dependent manner. Additionally, a significant increase of intracellular reactive oxygen species (ROS) was induced in A549 cells less than 0.5 h after DADS treatment, indicating that ROS may be an early event in DADS-modulated apoptosis. Treatment of A549 cells with N-acetyl cysteine (NAC) completely abrogated DADS-induced cell cycle arrest and apoptosis. The result indicated that oxidative stress modulates cell proliferation and cell death induced by DADS

  4. Nanostructured delivery system for zinc phthalocyanine: preparation, characterization, and phototoxicity study against human lung adenocarcinoma A549 cells

    Mariana da Volta Soares

    2011-01-01

    Full Text Available Mariana da Volta Soares1, Mainara Rangel Oliveira1, Elisabete Pereira dos Santos1, Lycia de Brito Gitirana2, Gleyce Moreno Barbosa3, Carla Holandino Quaresma3, Eduardo Ricci-Júnior11Department of Medicines, Laboratório de Desenvolvimento Galênico (LADEG, Faculty of Pharmacy, 2Laboratory of Animal and Comparative Histology, Glycobiology Research Program, Institute of Biomedical Science, 3Department of Medicines, Laboratório Multidisciplinar de Ciências Farmacêuticas, Faculty of Pharmacy, Federal University of Rio de Janeiro (UFRJ, Rio de Janeiro, BrazilAbstract: In this study, zinc phthalocyanine (ZnPc was loaded onto poly-ε-caprolactone (PCL nanoparticles (NPs using a solvent emulsification–evaporation method. The process yield and encapsulation efficiency were 74.2% ± 1.2% and 67.1% ± 0.9%, respectively. The NPs had a mean diameter of 187.4 ± 2.1 nm, narrow distribution size with a polydispersity index of 0.096 ± 0.004, zeta potential of -4.85 ± 0.21 mV, and spherical shape. ZnPc has sustained release, following Higuchi’s kinetics. The photobiological activity of the ZnPc-loaded NPs was evaluated on human lung adenocarcinoma A549 cells. Cells were incubated with free ZnPc or ZnPc-loaded NPs for 4 h and then washed with phosphate-buffered saline. Culture medium was added to the wells containing the cells. Finally, the cells were exposed to red light (660 nm with a light dose of 100 J/cm2. The cellular viability was determined after 24 h of incubation. ZnPc-loaded NPs and free photosensitizer eliminated about 95.9% ± 1.8% and 28.7% ± 2.2% of A549 cells, respectively. The phototoxicity was time dependent up to 4 h and concentration dependent at 0–5 µg ZnPc. The cells viability decreased with the increase of the light dose in the range of 10–100 J/cm2. Intense lysis was observed in the cells incubated with the ZnPc-loaded NPs and irradiated with red light. ZnPc-loaded PCL NPs are the release systems that promise photodynamic

  5. 氨甲蝶呤对映体获得性耐药A549细胞株二氢叶酸还原酶基因表达分析%Analysis for different expression of dihydrofolate reductase gene in methotrexate enantiomers-resistant lung cancer A549 cell lines

    李道静; 何晓东; 孙余婕; 凡任芝; 许维东; 孙利; 张永娟; 张白银; 沈佐君

    2011-01-01

    Objective To study the relationship between methotrexate (MTX) enantiomers resistance and levels of dihydrofolate reductase (DHFR) mRNA. Methods AS49 cells were exposed to intermittenfiy and progressively increasing dose of the two enantiomers of MTX. The expression of DHFR gene was assayed by real-time fluorescence quantitative polymerase chain reaction ( FQ-PCR ). Resuits The resistant indexes of cell lines were different for L-( +)-MTX and D-(-)-MTX enantiomer. D-(-)-MTX resistance cell lines showed higher resistant index than L-( + )-MTX resistant cell lines. The expression level of DHFR mRNA in the resistant cell lines was less than that of parent cells at the concentration of 15 μ mol/L of beth L-( + )- and D-(-)-MTX enantiomer (P > 0.05 ). The expression level of DHFR mRNA in resistant cell lines was relatively high at increasing concentration of 35 μmol/L and 45 μ mol/L of D-(-) MTX. The results of the FQ-PCR revealed that the MTX resistance was associated with increased expression of DHFR mRNA. Conclusion The expression of DHFR gene was inhibited after the cell lines induced by 15 μmol/L of D-(-) -MTX enantoimers in MTX resistant cell line. The expression of DHFR gene showed significant difference in chirality. DHFR mRNA should be examined during MTX treatment, which could be helpful to prognosticate the resistant status of cell line.%目的 研究氨甲蝶呤(MTX)对映体[L-(+)-MTX和D-(-)-MTX]耐药与二氢叶酸还原酶(DHFR)基因表达的关系.方法 用浓度递增结合低剂量持续诱导法获得A549细胞对不同构型及不同浓度的MTX对映体的耐药细胞株,荧光定量PCR检测耐药细胞株中DHFR基因的相对含量.结果 对两种不同对映体的获得性耐药存在差异,D型耐药细胞耐药指数高于L型;对映体各浓度耐药细胞间耐药指数也有差异.15 μmol/L L型、D型MTX首次诱导耐药细胞的DHFR相对含量低于亲本细胞,对该浓度对映体耐药的各细胞组间没有差别(P>0

  6. Characterization of indoor dust from Brazil and evaluation of the cytotoxicity in A549 lung cells.

    Deschamps, E; Weidler, P G; Friedrich, F; Weiss, C; Diabaté, S

    2014-04-01

    Over the past decade, ambient air particulate matter (PM) has been clearly associated with adverse health effects. In Brazil, small and poor communities are exposed to indoor dust derived from both natural sources, identified as blowing soil dust, and anthropogenic particles from mining activities. This study investigates the physicochemical and mineralogical composition of indoor PM10 dust samples collected in Minas Gerais, Brazil, and evaluates its cytotoxicity and inflammatory potential. The mean PM10 mass concentration was 206 μg/m(3). The high dust concentration in the interior of the residences is strongly related to blowing soil dust. The chemical and mineralogical compositions were determined by ICP-OES and XRD, and the most prominent minerals were clays, Fe-oxide, quartz, feldspars, Al(hydr)oxides, zeolites, and anatase, containing the transition metals Fe, Cr, V, Ni, Cu, Zn, Ti, and Mn as well as the metalloid As. The indoor dust samples presented a low water solubility of about 6 %. In vitro experiments were carried out with human lung alveolar carcinoma cells (A549) to study the toxicological effects. The influence of the PM10 dust samples on cell viability, intracellular formation of reactive oxygen species (ROS), and release of the pro-inflammatory cytokine IL-8 was analysed. The indoor dust showed little effects on alamarBlue reduction indicating unaltered mitochondrial activity. However, significant cell membrane damage, ROS production, and IL-8 release were detected in dependence of dose and time. This study will support the implementation of mitigation actions in the investigated area in Brazil. PMID:23990125

  7. Digoxin Downregulates NDRG1 and VEGF through the Inhibition of HIF-1α under Hypoxic Conditions in Human Lung Adenocarcinoma A549 Cells

    Dong Wei

    2013-04-01

    Full Text Available Digoxin, an inhibitor of Na+/K+ ATPase, has been used in the treatment of heart-related diseases (such as congestive heart failure and atrial arrhythmia for decades. Recently, it was reported that digoxin is also an effective HIF-1α inhibitor. We investigated whether digoxin could suppress tumor cell growth through HIF-1α in non-small cell lung cancer cells (A549 cells under hypoxic conditions. An MTT assay was used to measure cell viability. RT-PCR and western blotting were performed to analyze the mRNA and protein expression of VEGF, NDRG1, and HIF-1α. HIF-1α nuclear translocation was then determined by EMSA. Digoxin was found to inhibit the proliferation of A549 cells under hypoxic conditions. Our results showed that hypoxia led to the upregulation of VEGF, NDRG1, and HIF-1α both at the mRNA and protein levels. We also found that the hypoxia-induced overexpression of VEGF, NDRG1, and HIF-1α was suppressed by digoxin in a concentration-dependent manner. As expected, our EMSA results demonstrated that under hypoxic conditions HIF-1α nuclear translocation was also markedly reduced by digoxin in a concentration-dependent manner. Our results suggest that digoxin downregulated hypoxia-induced overexpression of VEGF and NDRG1 at the transcriptional level probably through the inhibition of HIF-1α synthesis in A549 cells.

  8. Multidimensional effects of biologically synthesized silver nanoparticles in Helicobacter pylori, Helicobacter felis, and human lung (L132) and lung carcinoma A549 cells

    Gurunathan, Sangiliyandi; Jeong, Jae-Kyo; Han, Jae Woong; Zhang, Xi-Feng; Park, Jung Hyun; Kim, Jin-Hoi

    2015-02-01

    Silver nanoparticles (AgNPs) are prominent group of nanomaterials and are recognized for their diverse applications in various health sectors. This study aimed to synthesize the AgNPs using the leaf extract of Artemisia princeps as a bio-reductant. Furthermore, we evaluated the multidimensional effect of the biologically synthesized AgNPs in Helicobacter pylori, Helicobacter felis, and human lung (L132) and lung carcinoma (A549) cells. UV-visible (UV-vis) spectroscopy confirmed the synthesis of AgNPs. X-ray diffraction (XRD) indicated that the AgNPs are specifically indexed to a crystal structure. The results from Fourier transform infrared spectroscopy (FTIR) indicate that biomolecules are involved in the synthesis and stabilization of AgNPs. Dynamic light scattering (DLS) studies showed the average size distribution of the particle between 10 and 40 nm, and transmission electron microscopy (TEM) confirmed that the AgNPs were significantly well separated and spherical with an average size of 20 nm. AgNPs caused dose-dependent decrease in cell viability and biofilm formation and increase in reactive oxygen species (ROS) generation and DNA fragmentation in H. pylori and H. felis. Furthermore, AgNPs induced mitochondrial-mediated apoptosis in A549 cells; conversely, AgNPs had no significant effects on L132 cells. The results from this study suggest that AgNPs could cause cell-specific apoptosis in mammalian cells. Our findings demonstrate that this environmentally friendly method for the synthesis of AgNPs and that the prepared AgNPs have multidimensional effects such as anti-bacterial and anti-biofilm activity against H. pylori and H. felis and also cytotoxic effects against human cancer cells. This report describes comprehensively the effects of AgNPs on bacteria and mammalian cells. We believe that biologically synthesized AgNPs will open a new avenue towards various biotechnological and biomedical applications in the near future.

  9. Effect of silencing of ATM expression by siRNA on radiosensitivity of human lung adenocarcinoma A549 cells

    Objective: To investigate the effect of silencing of ataxia-telangiectasia mutated (ATM) expression by plasmid-mediated RNA interference on the radiosensitivity of human lung adenocarcinoma A549 cells. Methods: Eukaryotic expression plasmid containing ATM small interfering RNA (siRNA) (pSilencer2.1-ATM), as well as pSilencer2.1-nonspecific, was constructed.Lung adenocarcinoma A549 cells were divided into positive group, negative group,and control group to be transfected with pSilencer2.1-ATM, pSilencer2.1-nonspecific, and no plasmid, respectively. The mRNA and protein expression of ATM was measured by RT-PCR and Western blot, respectively. The change in cell radiosensitivity was observed by colony-forming assay. Cell cycle and cell apoptosis were analyzed by flow cytometry. Results: The eukaryotic expression plasmid containing ATM siRNA was successfully constructed. The RT-PCR and Western blot demonstrated that the expression of ATM was down-regulated in the positive group. The sensitization enhancement ratios (D0 ratios) for the positive group and negative group were 1.50 and 1.01, respectively. The flow cytometry revealed that the proportions of A549 cells in G1 and G2/M phases were significantly lower in the positive group than in the control group (51.27% vs 61.85%, P = 0.012; 6.34% vs 10.91%, P = 0.008) and that the apoptosis rate was significantly higher in the positive group than in the control group and negative group (49.31% vs 13.58%, P = 0.000; 49.31% vs 13.17%, P = 0.000). Conclusions: Silencing of ATM expression may increase the radiosensitivity of human lung adenocarcinoma A549 cells, probably by affecting the cell cycle and promoting cell apoptosis. (authors)

  10. CK2 inhibitor CX-4945 blocks TGF-β1-induced epithelial-to-mesenchymal transition in A549 human lung adenocarcinoma cells.

    Jiyeon Kim

    Full Text Available BACKGROUND: The epithelial-to-mesenchymal transition (EMT is a major phenotype of cancer metastasis and invasion. As a druggable cancer target, the inhibition of protein kinase CK2 (formally named to casein kinase 2 has been suggested as a promising therapeutic strategy to treat EMT-controlled cancer metastasis. This study aimed to evaluate the effect of the CK2 inhibitor CX-4945 on the processes of cancer migration and invasion during the EMT in A549 human lung adenocarcinoma cells. MATERIALS AND METHODS: The effect of CX-4945 on TGF-β1-induced EMT was evaluated in A549 cells treated with TGF-β1 (5 ng/ml and CX-4945. The effect of CX-4945 on TGF-β1-induced cadherin switch and activation of key signaling molecules involved in Smad, non-Smad, Wnt and focal adhesion signaling pathways were investigated by Western blot analysis, immunocytochemistry and reporter assay. Additionally, the effect of CX-4945 on TGF-β1-induced migration and invasion was investigated by wound healing assay, Boyden chamber assay, gelatin zymography, and the quantitative real-time PCR. RESULTS: CX-4945 inhibits the TGF-β1-induced cadherin switch and the activation of key signaling molecules involved in Smad (Smad2/3, Twist and Snail, non-Smad (Akt and Erk, Wnt (β-catenin and focal adhesion signaling pathways (FAK, Src and paxillin that cooperatively regulate the overall process of EMT. As a result, CX-4945 inhibits the migration and invasion of A549 cells accompanied with the downregulation of MMP-2 and 9. CONCLUSIONS: Clinical evaluation of CX-4945 in humans as a single agent in solid tumors and multiple myeloma has established its promising pharmacokinetic, pharmacodynamic, and safety profiles. Beyond regression of tumor mass, CX-4945 may be advanced as a new therapy for cancer metastasis and EMT-related disorders.

  11. Activities of Ten Essential Oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 Cancer Cells

    Yuangang Zu

    2010-04-01

    Full Text Available Ten essential oils, namely, mint (Mentha spicata L.,Lamiaceae, ginger (Zingiber officinaleRosc.,Zingiberaceae, lemon (Citrus limon Burm.f.,Rutaceae, grapefruit (Citrus paradisi Macf., Rutaceae, jasmine (Jasminum grandiflora L.,Oleaceae, lavender (Mill.,Lamiaceae, chamomile (Matricaria chamomilla L., Compositae, thyme (Thymus vulgaris L., Lamiaceae, rose (Rosa damascena Mill.,Rosaceae and cinnamon (Cinnamomum zeylanicumN. Lauraceae were tested for their antibacterial activities towards Propionibacterium acnes and in vitro toxicology against three human cancer cell lines. Thyme, cinnamon and rose essential oils exhibited the best antibacterial activities towards P. acnes, with inhibition diameters of 40 ± 1.2 mm, 33.5 ± 1.5 mm and 16.5 ± 0.7 mm, and minimal inhibitory concentrations of 0.016% (v/v, 0.016% (v/v and 0.031% (v/v, respectively. Time-kill dynamic procedures showed that thyme, cinnamon, rose, and lavender essential oils exhibited the strongest bactericidal activities at a concentration of 0.25% (v/v, and P. acnes was completely killed after 5 min. The thyme essential oil exhibited the strongest cytotoxicity towards three human cancer cells. Its inhibition concentration 50% (IC50 values on PC-3, A549 and MCF-7 tumor cell lines were 0.010% (v/v, 0.011% (v/v and 0.030% (v/v, respectively. The cytotoxicity of 10 essential oils on human prostate carcinoma cell (PC-3 was significantly stronger than on human lung carcinoma (A549 and human breast cancer (MCF-7 cell lines.

  12. Activities of ten essential oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 cancer cells.

    Zu, Yuangang; Yu, Huimin; Liang, Lu; Fu, Yujie; Efferth, Thomas; Liu, Xia; Wu, Nan

    2010-05-01

    Ten essential oils, namely, mint (Mentha spicata L., Lamiaceae), ginger (Zingiber officinale Rosc., Zingiberaceae), lemon (Citrus limon Burm.f., Rutaceae), grapefruit (Citrus paradisi Macf., Rutaceae), jasmine (Jasminum grandiflora L., Oleaceae), lavender (Mill., Lamiaceae), chamomile (Matricaria chamomilla L., Compositae), thyme (Thymus vulgaris L., Lamiaceae), rose (Rosa damascena Mill., Rosaceae) and cinnamon (Cinnamomum zeylanicum N. Lauraceae) were tested for their antibacterial activities towards Propionibacterium acnes and in vitro toxicology against three human cancer cell lines. Thyme, cinnamon and rose essential oils exhibited the best antibacterial activities towards P. acnes, with inhibition diameters of 40 +/- 1.2 mm, 33.5 +/- 1.5 mm and 16.5 +/- 0.7 mm, and minimal inhibitory concentrations of 0.016% (v/v), 0.016% (v/v) and 0.031% (v/v), respectively. Time-kill dynamic procedures showed that thyme, cinnamon, rose, and lavender essential oils exhibited the strongest bactericidal activities at a concentration of 0.25% (v/v), and P. acnes was completely killed after 5 min. The thyme essential oil exhibited the strongest cytotoxicity towards three human cancer cells. Its inhibition concentration 50% (IC(50)) values on PC-3, A549 and MCF-7 tumor cell lines were 0.010% (v/v), 0.011% (v/v) and 0.030% (v/v), respectively. The cytotoxicity of 10 essential oils on human prostate carcinoma cell (PC-3) was significantly stronger than on human lung carcinoma (A549) and human breast cancer (MCF-7) cell lines. PMID:20657472

  13. Differential Regulation of Gene Expression of Alveolar Epithelial Cell Markers in Human Lung Adenocarcinoma-Derived A549 Clones

    Hiroshi Kondo

    2015-01-01

    Full Text Available Stem cell therapy appears to be promising for restoring damaged or irreparable lung tissue. However, establishing a simple and reproducible protocol for preparing lung progenitor populations is difficult because the molecular basis for alveolar epithelial cell differentiation is not fully understood. We investigated an in vitro system to analyze the regulatory mechanisms of alveolus-specific gene expression using a human alveolar epithelial type II (ATII cell line, A549. After cloning A549 subpopulations, each clone was classified into five groups according to cell morphology and marker gene expression. Two clones (B7 and H12 were further analyzed. Under serum-free culture conditions, surfactant protein C (SPC, an ATII marker, was upregulated in both H12 and B7. Aquaporin 5 (AQP5, an ATI marker, was upregulated in H12 and significantly induced in B7. When the RAS/MAPK pathway was inhibited, SPC and thyroid transcription factor-1 (TTF-1 expression levels were enhanced. After treatment with dexamethasone (DEX, 8-bromoadenosine 3′5′-cyclic monophosphate (8-Br-cAMP, 3-isobutyl-1-methylxanthine (IBMX, and keratinocyte growth factor (KGF, surfactant protein B and TTF-1 expression levels were enhanced. We found that A549-derived clones have plasticity in gene expression of alveolar epithelial differentiation markers and could be useful in studying ATII maintenance and differentiation.

  14. Lung Cancer

    Lung cancer is one of the most common cancers in the world. It is a leading cause of cancer death in men and women in the United States. Cigarette smoking causes most lung cancers. The more cigarettes you smoke per day and ...

  15. What Is Lung Cancer?

    ... starts in the lungs, it is called lung cancer. Lung cancer begins in the lungs and may spread ... lung cancer. For more information, visit the National Cancer Institute’s Lung Cancer. Previous Basic Information Basic Information Basic Information ...

  16. Immune Evasion of Human Lung Carcinoma Cell A549 Suppressed by Human Lymphoma Cell Jurkat via Fas/FasL Pathway

    Hongmei WANG

    2010-07-01

    Full Text Available Background and objective Tumor escape from the host immune system has been a major problem in immunotherapy of human malignancies. FasL/Fas-induced apoptosis plays an important role in various immunological processes. The aim of this study is to investigate the immune evasion in human lung carcinoma cell A549 induced by human lymphoma cell Jurkat via Fas/FasL pathway. Methods Jurkat cells and A549 cells were co-cultured at different proportions. The apoptotic rates of A549 cells were determined by flow cytometry (FCM. Protein levels of Fas, FasL and Caspase-8 in A549 cells were detected by Western blot. Results Survival rates of A549 cells gradually decreased and apoptotic rates of A549 cells were significantly enhanced along with ratio increasing between Jurkat and A549. Meanwhile, the protein levels of Fas and Caspase-8 gradually increased in A549 cells, and the protein levels of FasL had no significant difference in all groups. Conclusion The Jurkat cells might decrease the survival rates of A549 cells and enhanced its apoptosis and immune evasion partly via Fas/FasL pathway.

  17. Lung Cancer Prevention

    ... Treatment Lung Cancer Prevention Lung Cancer Screening Research Lung Cancer Prevention (PDQ®)–Patient Version What is prevention? Go ... to keep cancer from starting. General Information About Lung Cancer Key Points Lung cancer is a disease in ...

  18. Effects of paclitaxel on cell proliferation and apoptosis and its mechanism in human lung adenocarcinoma A549 cells

    Baoan Gao; Chunling Du; Wenbo Ding; Shixiong Chen; Jun Yang

    2006-01-01

    Objective: To investigate the effect of paclitaxel on cell proliferation and apoptosis of human lung adenocarcinoma A549 cells line and its mechanism in vitro. Methods: Cell growth inhibition of paclitaxel on A549 cells was analyzed by MTT assay. Cell apoptosis was detected by DNA cytofluorometry, Hoechst33258 staining when treated with paclitaxel for 48hours. Meanwhile, Cell cycle and apoptotic rate were analyzed by flow cytometry. The protein expressions of Bax and Bcl-2 were studied by Western Blot. Results: Paclitaxel inhibited the proliferation of A549 cells in a time-and dose-dependant manner.Hoechst33258 staining indicated that apoptosis was induced by paclitaxel. After treated for 48 hours, cell apoptosis rates of 25nmol/L, 50 nmol/L and 100 nmol/L paclitaxel groups were 11.52 ± 1.94% ,17.73 ± 2.53%, and 29.32 ± 5.51% respectively,which were significantly higher than those of control group 5.88 ± 1.07%(all P < 0.01 ), and apoptosis rate increased in dose-dependant manner. Meanwhile, G2/M stage cell percentage of 25 nmol/L, 50 nmol/L and 100 nmol/L paclitaxel groups were 42.52± 6.25%, 40.46 ± 5.81%, and 35.34 ± 6.17% respectively,which were significantly higher than that of control group 22.32 ±3.30%(all P < 0.01 ); Western blot showed that paclitaxel increased the expression of Bax and decreased the expression of Bcl-2 in dose-dependant manner. Conclusion: Paclitaxel can inhibit A549 cell proliferation in a time- and dose-dependant manner. Its mechanism may be related to arresting cell cycle in G2/M stage and induce cell apoptosis by up-modulating Bax expression and down-modulating Bcl-2 expression.

  19. Factors influencing recovery from potentially lethal radiation damage in A549 human lung carcinoma cells

    Plateau phase A549 cells exhibit potentially lethal radiation damage recovery (PLDR) that is dependent upon both the pH and the glucose content of the spent medium. At 9-10 days after plating, unfed A549 plateau cultures are acidic (pH 6.5 - 6.7), contain 2 to 4 mM glucose, and exhibit an approximately 40-fold increase in survival when held for 6 hrs in spent medium vs being subcultured immediately after 10 Gy aerobic irradiation. PLDR is maximal 24 hrs. post-irradiation. Adjustment of the pH of the spent medium to 7.5, by NaOH addition, either prior to or immediately post irradiation, nearly completely inhibits PLDR in this cell line. The authors found that medium acidity inhibits glucose utilization, and that alkalinization of spent medium, to pH 7.5, results in stimulation of glucose consumption. Plateau phase cultures depleted of glucose, as a result of medium alkalinization, are not capable of PLDR. In addition to pH effects, they observed that several agents, including nicotinamide, 3-aminobenzamide, caffeine, 2-deoxyglucose and glucosamine, partially inhibit PLDR in A549 plateau phase cultures

  20. Runx3 Expression Inhibits Proliferation and Distinctly Alters mRNA Expression of Bax in AGS and A549 Cancer Cells

    Torshabi, Maryam; Faramarzi, Mohammad Ali; Tabatabaei Yazdi, Mojtaba; Ostad, Seyyed Naser; Gharemani, Mohammad Hosein

    2011-01-01

    Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue–restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS and A549 cells using fugene6 and PolyFect and Runx3 expression was confirmed by fluorescent microscopy and RT-PCR. The effect of Runx3 transfection on cell proliferation was determined by MTT assay and the results were confirmed by the trypan blue dye exclusion method. The effect of Runx3 expression on mRNA expression of BCL2-associated X protein (Bax) was evaluated using RT-PCR. In AGS and A549 cells, Runx3 expression inhibited cell proliferation (p < 0.01). The growth inhibition was less in A549 cells. We show that Runx3 expression increases Bax mRNA expression in AGS cells when compared with control (p < 0.05), but no significant differences in mRNA expression was observed in both examined cells. Runx3 expression has antiproliferative effect in AGS cell perhaps via increase in expression of Bax. The effect of Runx3 on A549 cells’ viability which has endogenous level of Runx3 is not related to Bax. These findings implicate a complex regulation by Runx3 in inhibition of cell proliferation utilizing Bax. PMID:24250365

  1. Lung Cancer Screening

    ... Treatment Lung Cancer Prevention Lung Cancer Screening Research Lung Cancer Screening (PDQ®)–Patient Version What is screening? Go ... These are called diagnostic tests . General Information About Lung Cancer Key Points Lung cancer is a disease in ...

  2. Cytochrome c oxidase is activated by the oncoprotein Ras and is required for A549 lung adenocarcinoma growth

    Telang Sucheta

    2012-08-01

    Full Text Available Abstract Background Constitutive activation of Ras in immortalized bronchial epithelial cells increases electron transport chain activity, oxygen consumption and tricarboxylic acid cycling through unknown mechanisms. We hypothesized that members of the Ras family may stimulate respiration by enhancing the expression of the Vb regulatory subunit of cytochrome c oxidase (COX. Results We found that the introduction of activated H-RasV12 into immortalized human bronchial epithelial cells increased eIF4E-dependent COX Vb protein expression simultaneously with an increase in COX activity and oxygen consumption. In support of the regulation of COX Vb expression by the Ras family, we also found that selective siRNA-mediated inhibition of K-Ras expression in A549 lung adenocarcinoma cells reduced COX Vb protein expression, COX activity, oxygen consumption and the steady-state concentration of ATP. We postulated that COX Vb-mediated activation of COX activity may be required for the anchorage-independent growth of A549 cells as soft agar colonies or as lung xenografts. We transfected the A549 cells with COX Vb small interfering or shRNA and observed a significant reduction of their COX activity, oxygen consumption, ATP and ability to grow in soft agar and as poorly differentiated tumors in athymic mice. Conclusion Taken together, our findings indicate that the activation of Ras increases COX activity and mitochondrial respiration in part via up-regulation of COX Vb and that this regulatory subunit of COX may have utility as a Ras effector target for the development of anti-neoplastic agents.

  3. Enhancement of radiosensitivity by topoisomerase II inhibitor, amrubicin and amrubicinol, in human lung adenocarcinoma A549 cells and kinetics of apoptosis and necrosis induction

    Hayashi, Sachiko; Hatashita, Masanori; Matsumoto, Hideki; Shioura, Hiroki; KITAI, Ryuhei; Kano, Eiichi

    2006-01-01

    The effects of amrubicin (AMR) and its activemetabolite, amrubicinol (AMROH), on the sensitivity ofhuman lung adenocarcinoma A549 cells to ionizing radiationwere investigated in vitro. Further, the kinetics of apoptosisand necrosis induction were also analyzed. The cytocidalefftcts of X-ray irradiation on A549 cells resulted in a lowlevel of radiosensitivity with a D value of 12 Gy. The slopesof the survival curves in the exponential phase were plottedon semilogarithmic paper for radiation co...

  4. Α-MMC and MAP30, two ribosome-inactivating proteins extracted from Momordica charantia, induce cell cycle arrest and apoptosis in A549 human lung carcinoma cells.

    Fan, Xiang; He, Lingli; Meng, Yao; Li, Gangrui; Li, Linli; Meng, Yanfa

    2015-05-01

    α‑Momorcharin (α‑MMC) and momordica anti‑human immunodeficiency virus protein (MAP30), produced by Momordica charantia, are ribosome‑inactivating proteins, which have been reported to exert inhibitory effects on cultured tumor cells. In order to further elucidate the functions of these agents, the present study aimed to investigate the effects of α‑MMC and MAP30 on cell viability, the induction of apoptosis, cell cycle arrest, DNA integrity and superoxide dismutase (SOD) activity. α‑MMC and MAP30 were purified from bitter melon seeds using ammonium sulfate precipitation in combination with sulfopropyl (SP)‑sepharose fast flow, sephacryl S‑100 and macro‑Cap‑SP chromatography. MTT, flow cytometric and DNA fragmentation analyses were then used to determine the effects of α‑MMC and MAP30 on human lung adenocarcinoma epithelial A549 cells. The results revealed that A549 cells were sensitive to α‑MMC and MAP30 cytotoxicity assays in vitro. Cell proliferation was significantly suppressed following α‑MMC and MAP30 treatment in a dose‑ and time‑dependent manner; in addition, the results indicated that MAP30 had a more potent anti‑tumor activity compared with that of α‑MMC. Cell cycle arrest in S phase and a significantly increased apoptotic rate were observed following treatment with α‑MMC and MAP30. Furthermore, DNA integrity analysis revealed that the DNA of A549 cells was degraded following treatment with α‑MMC and MAP30 for 48 h. The pyrogallol autoxidation method and nitrotetrazolium blue chloride staining were used to determine SOD activity, the results of which indicated that α‑MMC and MAP30 did not possess SOD activity. In conclusion, the results of the present study indicated that α‑MMC and MAP30 may have potential as novel therapeutic agents for the prophylaxis and treatment of cancer. PMID:25573293

  5. Growth arrest of lung carcinoma cells (A549) by polyacrylate-anchored peroxovanadate by activating Rac1-NADPH oxidase signalling axis.

    Chatterjee, Nirupama; Anwar, Tarique; Islam, Nashreen S; Ramasarma, T; Ramakrishna, Gayatri

    2016-09-01

    Hydrogen peroxide is often required in sublethal, millimolar concentrations to show its oxidant effects on cells in culture as it is easily destroyed by cellular catalase. Previously, we had shown that diperoxovanadate, a physiologically stable peroxovanadium compound, can substitute H2O2 effectively in peroxidation reactions. We report here that peroxovanadate when anchored to polyacrylic acid (PAPV) becomes a highly potent inhibitor of growth of lung carcinoma cells (A549). The early events associated with PAPV treatment included cytoskeletal modifications, increase in GTPase activity of Rac1, accumulation of the reactive oxygen species, and also increase in phosphorylation of H2AX (γH2AX), a marker of DNA damage. These effects persisted even at 24 h after removal of the compound and culminated in increased levels of p53 and p21 together with growth arrest. The PAPV-mediated growth arrest was significantly abrogated in cells pre-treated with the N-acetylcysteine, Rac1 knocked down by siRNA and DPI an inhibitor of NADPH oxidase. In conclusion, our results show that polyacrylate derivative of peroxovanadate efficiently arrests growth of A549 cancerous cells by activating the axis of Rac1-NADPH oxidase leading to oxidative stress and DNA damage. PMID:27435854

  6. Lung cancer

    There will be over 160,000 cases of lung cancer diagnosed in the US in 1991, and deaths from this disease account for a quarter of all cancer deaths in this country. The incidence of lung cancer has continued to increase, especially among women. With 31% of American men and 25% of American women identified in the 1985 census as cigarette smokers, it is likely that this trend will continue well into the next century. Unfortunately, the majority of patients present with locally advanced tumors or distant metastatic disease. Presently, most patients with lung cancer will receive radiation therapy either in an attempt to control inoperable or locally advanced disease, or for palliation of symptomatic intrathoracic or metastatic disease. Because of the poor prognosis of all patients excepting those with early stage resectable lesions, lung cancer is appropriately the subject of intense clinical investigation and controversy throughout the world

  7. Effect of RNAi targeting survivin gene combined with X-rays radiation on apoptosis of lung adenocarcinoma A549 cells

    Objective: To construct the vector of RNA interference (RNAi) targeting survivin gene and observe its effect combined with X-rays radiation on lung adenocarcinoma A549 cell apoptosis. Methods: One pair of RNAi sequence targeting survivin gene were designed according to its cDNA sequence reported in GenBank, the recombinant RNAi plasmid pGenesil2-survivin was constructed. After identified by enzyme digestion and sequencing, the pGenesil2-survivin plasmid was trasfeced into A549 cells.In the experiment, normal group,pGenesil2 group, pGenesil2-survivin group,5 Gy irradiation group and pGenesil2-survivin + 5 Gy irradiation group were set up.The apoptosis of A549 cells was measured by flow cytometry with PI/Annexin V and TUNEL,the survivin and caspase-3 expressions were measured by Western blotting. Results: Two fragments about 389 bp and 4 206 bp were gotten by Kpn I and EcoR I enzyme digestion, they are the same to expected result, the sequencing result was compared to oligonucleotide chain with DNAssist 2.0, they were equal, these indicated the identification of pGenesil2-survivin vector was right; pGenesil2-survivin was transfected into A549 cells for 48 h, the apoptotic percentage in pGenesil2-survivin and 5 Gy X-rays groups increased obviously (P< 0.05), when the both were combined, the effect was more obvious;the Western blotting results appeared that the survivin gray scale/β-actin gray scale in pGenesil2-survivin group was lower than that in normal group(P< 0.01), and the caspase-3 gray scale/β-actin gray scale was higher than that in normal group,and that ratio in pGenesil2-survivin+5 Gy irradiation group was more high(P< 0.01). Conclusion: RNAi targeting surviving gene could inhibit survivin protein expression,but enhance caspase-3 protein expression, and promote apoptosis. When it is combined with 5 Gy X-rays irradiation, the promotion of apoptosis is enhanced. (authors)

  8. X射线对人肺腺癌A549细胞Pokemon基因表达的影响%Effects of X-ray irradiation on expression of Pokemon gene in A549 cells of human lung adenocarcinoma

    王璐; 邹跃; 江其生; 李伟; 宋秀军; 周湘艳; 王翠兰

    2011-01-01

    目的 研究不同剂量X射线照射及照射后不同时间点对人肺腺癌A549细胞Pokemon基因表达的影响.方法 用吸收剂量分别为2、4、6和8 Gy的X射线照射体外堵养的人肺腺癌A549细胞,2、4、8、12、24、48和72 ha,用实时定量PCR技术检测其中的Pokemon mRNA表达水平,以未照射组为对照.结果 在2、4、6、8 Gy X射线照射后的早期(除2 Gy照射后的2和4 h外)Pokemon mRNA的表达降低,但在晚期(48 h以后)呈升高趋势,在大部分时间点实验组与对照组的差异有统计学意义(t=3.40~154.76,P<0.05).结论 较大剂量的X射线在早期可下调A549细胞Pokemon基因mRNA的表达,诱导肿瘤细胞凋亡;但在晚期又可诱导A549细胞高表达PokemonmRNA,这可能与辐射所致A549细胞的DNA损伤修复和细胞周期调控有关.%Objective To study the dose-and time-effects of X-ray irradiation on the expression of Pokemon gene in A549 cells of human lung adenocarcinoma.Methods A549 cells were cultured in vitro and exposed to X-rays with the doses of 2,4,6 and 8 Gy,respectively.Untreated A549 cells were used as control group.The relative levels of Pokemon mRNA expression in the cells were detected by using quantitative real-time PCR at 2,4,8,12,24,48 and 72 h after irradiation.Results The Pokemon mRNA expression levels decreased in the early period after irradiation(except 2 and 4 h after irradiation in 2 Gy group)and then increased in the later stage(48 h after irradiation)with significant statistical differences at the most time points in comparison with the control group(t=3.40-154.76,P<0.05).Conclusions Higher doses of X-rays may degrade the expression of Pokemon mRNA in the human A549 cells and induce apoptosis in the early period,hut also may upgrade its expression in the later period, which might be correlated with the cell cycle regulation and DNA damage repair in the A549 cells.

  9. 甲基化抑制剂5-氮杂2′-脱氧胞苷对三维培养A549细胞辐射敏感性的影响%Effect of DNA Methyltransferase Inhibitor 5-aza-2′-deoxycytidine on Radiosensitivity of the Human Lung Cancer Cells in Three-dimensional Culture

    潘冬; 陈亚雄; 薛刚; 李小满; 任振新; 杜亚蓉; 胡步荣

    2014-01-01

    . Pretreatment with 5-Aza-CdR inhibited the A549 cell proliferation significantly. More micronucleus were observed after irradiation in 3D cells pretreated with 2 and 5 µmol/L concentration of drug than those without treatment. The survival fractions of cells pretreated by both 2 and 5 µmol/L drug reduced significantly in 3D cultures after irradiation. These significances, however, were found in 2D cells pretreated by only 5 µmol/L drug. Our results suggest that 5-Aza-CdR can inhibit the A549 cells proliferation and apparently enhance the radiosensitivity of cells in 3D cultures. Using of the low dose 5-Aza-CdR in clinical radiotherapy may reduce side effects and enhance effectively the cancer target therapy.

  10. A Novel Bioluminescence Orthotopic Mouse Model for Advanced Lung Cancer

    Li, Bo; Torossian, Artour; Li, Wenyan; Schleicher, Stephen; Niu, Kathy; Giacalone, Nicholas J; Kim, Sung June; Chen, Heidi; Gonzalez, Adriana; Moretti, Luigi; Lu, Bo

    2011-01-01

    Lung cancer is the leading cause of cancer-related death in the United States despite recent advances in our understanding of this challenging disease. An animal model for high-throughput screening of therapeutic agents for advanced lung cancer could help promote the development of more successful treatment interventions. To develop our orthotopic lung cancer model, luciferase-expressing A549 cancer cells were injected into the mediastinum of athymic nude mice. To determine whether the model ...

  11. Comparison of the uptakes of Tc-99m MIBI and Tc-99m tetrofosmin in A549, an MRP-expressing cancer cell, in vitro and in vivo

    Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Western blot analysis and immunohistochemistry were used for detetion of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at 100 μM of verapamil (Vrp), 50 μM of cyclosporin A (CsA) and 25 μM of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 60 min at 37.deg.C, using single cell suspensions at 1x106 cells/ml. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetorfosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at

  12. Comparison of the uptakes of Tc-99m MIBI and Tc-99m tetrofosmin in A549, an MRP-expressing cancer cell, in vitro and in vivo

    Yoo, Jeong Ah; Jeong, Shin Young; Seo, Myung Rang; Bae, Jin Ho; Ahn, Byeong Cheol; Lee, Kyu Bo; Lee, Jae Tae [School of Medicine, Kyungpook National Univ., Daegu (Korea, Republic of); Choi, Sang Woon; Lee, Byung Ho [Korea Institute of Chemical Technology, Daejon (Korea, Republic of)

    2003-12-01

    Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Western blot analysis and immunohistochemistry were used for detetion of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at 100 {mu}M of verapamil (Vrp), 50 {mu}M of cyclosporin A (CsA) and 25 {mu}M of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 60 min at 37.deg.C, using single cell suspensions at 1x10{sup 6} cells/ml. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetorfosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60

  13. Effect of glutathione depletion on the aerobic radiation response of A549 human lung carcinoma cells

    The authors demonstrated that depletion of glutathione (GSH) from cultured A549 cells to non-detectable levels, using L-buthionine sulfoximine (L-BSO), results in an increased aerobic radiation response. This response can be further increased if dimethylfumarate (DMF) is added concurrently with L-BSO. L-BSO is a relatively slow depletor of GSH compared to DMF, which acts by both spontaneous and enzyme catalysed reactions. The authors have studied: 1. the effect of continuous long-term exposure to 0.1 mM L-BSO on GSH levels and the subsequent radiation response and 2. the effect of GSH depletion on enzymes essential for radical detoxification. The results show an enhanced aerobic radiation response that increases with the time of exposure to L-BSO. For example surviving fraction (S.F.) after 5 Gy for cells exposed to L-BSO for 24 hrs is 0.004 and 0.08 for control cultures. Cells washed free of medium and irradiated in Hanks' show 0.0007 S.F. after 120 hr exposure to L-BSO and S.F. of 0.075 for the control cultures. The relationship between the chronic GSH depleted state, GSH peroxidase, and radiation induced lipid peroxidation is being investigated

  14. THE EFFECT OF IRISQUINONE ON THE GLUTATHIONE SYSTEM AND MRP EXPRESSION OF CISPLATIN-RESISTANT HUMAN LUNG ADENOCARCINOMA CELL LINE (A549DDP)

    LIANG; li

    2001-01-01

    [1] Li DH. A novel radiosensitizer "ANKA" for tumor (Irisquinone) [J]. Chin J Clin Oncol 1999; 26:153.[2]Bordow SB, Haber M, Madafiglio J, et al. Expression of the multidrug resistance-associated protein (MRP) gene correlates with amplification and overexpression of the N-myc oncogene in childhood neuroblastoma [J]. Cancer Res 1994; 54:5036.[3]Cai P, Liu XY, Han FS, et al. Establishment human lung adenocarcinoma cisplatin-resistant cell line A549DDP and the mechanism of its drug resistance [J]. Chin J Clin Oncol 1995; 22:582.[4]Cai P, Liu XY, Wang P. The value of glutathione reductase recycling assay measurement of content of glutathione in human plasma during tumor chemotherapy [J]. Chin J Clin Oncol l994; 21:717.[5]Zhan MC, Liu XY, Cai P, et al. Mechanism of resistance of human cell line A549DDP to cisplatin [J]. Chin J Clin Oncol 1998; 25:726.[6]Wang J, Liu XY, Wu MN, et al. Expression and reversion of drug resistance- and apoptosis- related genes of a DDP-resistant lung adeno-carcinoma cell line A549DDP [J]. Chin J Oncol 1999; 21:422.[7]Ishikawa T. The ATP-dependent glutathione S-conjugate export pump [J]. Treads Biol Sci 1992; 17:463.[8]Goto S, Yoshida K, Morikawa T, et al. Augmen-tation of transport for cisplatin-glutathione adduct in cisplatin-resistant cancer cells [J]. Cancer Res 1995; 55:4297.[9]Fujil R, Mutoh M, Sumizama T, et al. Adenosine triphosphate-dependent transport of leukotriene C4 by membrane vesicles prepared from cis-platinum-resistant human epidermoid carcinoma tumor cells [J]. JNCI 1994; 86:1781.[10]Ishikawa T, Ali-Osman F. Glutathion-associated cis-diamminedichloroplatinum (II) metabolism and ATP-dependent efflux from leukemia cells [J]. J Biol Chem 1993; 268:20116.[11]Ishikawa T, Wrighe CE, Ishizuka H. GS-X pumq is function ally overexpressed in cis-diammine-dichloroplatinum (II)-resistant human leukemia HL-60 cells and downregulated by cell differentiation [J]. J Biol Chem 1994; 269: 29085.

  15. Lung cancer - small cell

    Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC are ...

  16. Lung cancer - small cell

    Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC ...

  17. 三种缝线材料对人肺腺癌细胞A549增殖和细胞周期的影响%Effect of three suture lines on the proliferation and cell cycle of lung adenocarcinoma cell A549 in vitro

    Lianhua Ye; Yunchao Huang; Qilin Jin; Feng Hua; Guangqiang Zhao

    2011-01-01

    Objective: The interaction of cell and medical biomaterial is one of the significant factors to affect clinical application of medical biomaterial. This research is to investigate three of suture lines how to affect the proliferation and cell cycle of lung adenocarcinoma cell A549 in vitro. Methods: Three of suture lines were respectively cultivated with lung adenocarcinoma cell A549, after of 72 hours, we detected absorptions of each group by MTT method in order to reflect the proliferation of lung adenocarcinoma cell A549, and also examined percentage of G1 period cells and S period cells of each group by flow cytometry. Results: Different of suture lines had different effects on the proliferation and cell cycle of lung adenocarcinoma cell A549 (P < 0.05). The effect of absorbent suture line was the strongest on the proliferation and cell cycle of lung adenocarcinoma cell A549, the effect of chorda serica chirurgicalis was medium, and the effect of slide wire was poor. Different length of each suture line had different effects on the proliferation and cell cycle of lung adenocarcinoma cell A549 (P < 0.05).Conclusion: Three of suture line materials have different effects on the proliferation and cell cycle of lung adenocarcinoma cell A549, with dose-effect relationship.

  18. Effects of gene F10 over-expression on the tumorigenicity of A549 cells

    Ya-li SONG

    2012-07-01

    expressions of Bax and Caspase-3 were weak in F10+A549 group. Conclusion F10 gene may down-regulate expressions of Caspase-3 and Bax, and up-regulate expression of Bcl-2, which further enhance the tumorigenicity of lung cancer cell line A549 in nude mice.

  19. Nanostructured delivery system for zinc phthalocyanine: preparation, characterization, and phototoxicity study against human lung adenocarcinoma A549 cells

    da Volta Soares, Mariana; Oliveira, Mainara Rangel; dos Santos, Elisabete Pereira; de Brito Gitirana, Lycia; Barbosa, Gleyce Moreno; Quaresma, Carla Holandino; Ricci-Júnior, Eduardo

    2011-01-01

    In this study, zinc phthalocyanine (ZnPc) was loaded onto poly-ɛ-caprolactone (PCL) nanoparticles (NPs) using a solvent emulsification–evaporation method. The process yield and encapsulation efficiency were 74.2% ± 1.2% and 67.1% ± 0.9%, respectively. The NPs had a mean diameter of 187.4 ± 2.1 nm, narrow distribution size with a polydispersity index of 0.096 ± 0.004, zeta potential of −4.85 ± 0.21 mV, and spherical shape. ZnPc has sustained release, following Higuchi’s kinetics. The photobiological activity of the ZnPc-loaded NPs was evaluated on human lung adenocarcinoma A549 cells. Cells were incubated with free ZnPc or ZnPc-loaded NPs for 4 h and then washed with phosphate-buffered saline. Culture medium was added to the wells containing the cells. Finally, the cells were exposed to red light (660 nm) with a light dose of 100 J/cm2. The cellular viability was determined after 24 h of incubation. ZnPc-loaded NPs and free photosensitizer eliminated about 95.9% ± 1.8% and 28.7% ± 2.2% of A549 cells, respectively. The phototoxicity was time dependent up to 4 h and concentration dependent at 0–5 μg ZnPc. The cells viability decreased with the increase of the light dose in the range of 10–100 J/cm2. Intense lysis was observed in the cells incubated with the ZnPcloaded NPs and irradiated with red light. ZnPc-loaded PCL NPs are the release systems that promise photodynamic therapy use. PMID:21499420

  20. Radio-sensitization effect of an mTOR inhibitor, temsirolimus, on lung adenocarcinoma A549 cells under normoxic and hypoxic conditions

    The mammalian target of rapamycin (mTOR) correlates with cell survival under hypoxia and regulates hypoxia-inducible factor-1α (HIF-1α), a key protein in hypoxia-related events. However, the role of mTOR in radio-resistance has not been fully investigated. Therefore, the effect of mTOR on the radio-resistance of cancer cells under hypoxia was evaluated using the mTOR inhibitor temsirolimus. Clonogenic survival was examined in the A549 human lung adenocarcinoma cell line under normoxia or hypoxia, with or without temsirolimus. An oxygen enhancement ratio (OER) was calculated using the D10 values, the doses giving 10% survival. Western blotting was performed to investigate the effect of temsirolimus on mTOR and the HIF-1α pathway under normoxia and hypoxia. A549 cells showed a radio-resistance of 5.1 and 14.2 Gy, as indicated by D10 values under normoxia and hypoxia, respectively; the OER was 2.8. The cell survival rates under hypoxia and with temsirolimus remarkably decreased compared with those under normoxia. The D10 values of the cells under normoxia and hypoxia were 4.8 and 5.4 Gy, respectively (OER = 1.1). mTOR expression was suppressed by temsirolimus under both normoxia and hypoxia. HIF-1α expression decreased under hypoxia in the presence of temsirolimus. These results suggest that temsirolimus can overcome the radio-resistance induced by hypoxia. When the fact that mTOR acts upstream of HIF-1α is considered, our data suggest that the restoration of radiation sensitivity by temsirolimus under hypoxia may be associated with the suppression of the HIF-1α pathway. Temsirolimus could therefore be used as a hypoxic cell radio-sensitizer. (author)

  1. 6 Common Cancers - Lung Cancer

    ... Home Current Issue Past Issues 6 Common Cancers - Lung Cancer Past Issues / Spring 2007 Table of Contents For ... for Desperate Housewives. (Photo ©2005 Kathy Hutchins / Hutchins) Lung Cancer Lung cancer causes more deaths than the next ...

  2. 6 Common Cancers - Lung Cancer

    ... Bar Home Current Issue Past Issues 6 Common Cancers - Lung Cancer Past Issues / Spring 2007 Table of Contents ... Desperate Housewives. (Photo ©2005 Kathy Hutchins / Hutchins) Lung Cancer Lung cancer causes more deaths than the next three ...

  3. Lung cancer

    This article is about the diagnosis, treatment and monitoring of lung cancer. Before the treatment the histological samples allowing the diagnosis as well as its histological variety. The diagnosis include techniques such as bronchoscopy, ultrasound, tomography, puncture and endoscopic thoracotomy. The chemotherapy and radiotherapy are the main techniques used for the treatment

  4. Lung cancer

    Hansen, H H; Rørth, M

    1999-01-01

    The results of the many clinical trials published in 1997 had only modest impact on the treatment results using either cytostatic agents alone or combined with radiotherapy in lung cancer. In SCLC, combination chemotherapy including platin-compounds (cisplatin, carboplatin) and the podophyllotoxins...

  5. 骨髓间充质干细胞参与A549肺腺癌的组织修复★%Bone marrow mesenchymal stem cells are involved in tissue repair of A549 lung adenocarcinoma

    许峰; 张雷; 潘晋坤; 薛利利; 赵晓燕; 李宝平

    2013-01-01

    BACKGROUND:Tumor has been considered as a specific nonhealing trauma. Bone marrow mesenchymal stem cel s participate in tumor mesenchymal reconstitution by tumor tissue homing and differentiation into mesenchyme, resulting in changing tumor microenvironment and affecting tumor growth and transfer. OBJECTIVE:To explore the mechanisms of participation of bone marrow mesenchymal stem cel s in tumor tissue repair in an A549 lung cancer-bearing mouse model. METHODS:Bone marrow mesenchymal stem cel s were isolated in vitro, cultured, and identified using flow cytometry, and then used to establish a mouse model of A549 lung cancer-bearing. In the experimental group, human bone marrow mesenchymal stem cel s were injected into tissue surrounding the tumor. In the control group, an equal volume of PBS was injected. Animal survival condition and tumor size were compared. At 4 weeks, the specimens were harvested. Hematoxylin-eosin staining was used to compare tumor tissue. Masson staining was utilized to compare col agen fiber content. Reverse transcription-PCR was employed to detect the expression ofα-smooth muscle actin. Immunohistochemistry was used to examine the expression of fibroblast specific protein and fibroblast activation protein to reflect the degree of interstitial fibers in tumor tissue in both groups. The expression levels of vascular endothelial growth factor, hepatocyte growth factor, interleukin-6 and tenescin-C were compared between the two groups using immunohistochemistry. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s promoted tumor growth in tumor-bearing mice. The growth rate of tumor tissue in experimental group was faster than the control group (P<0.05). Compared with the control group,α-smooth muscle actin mRNA expression was significantly higher in the experimental group. Immunohistochemistry was used to detect the expression of tumor angiogenesis factors markers (fibroblast specific protein and fibroblast activation protein) in tumor

  6. Novel functional view of the crocidolite asbestos-treated A549 human lung epithelial transcriptome reveals an intricate network of pathways with opposing functions

    Stevens John R

    2008-08-01

    Full Text Available Abstract Background Although exposure to asbestos is now regulated, patients continue to be diagnosed with mesothelioma, asbestosis, fibrosis and lung carcinoma because of the long latent period between exposure and clinical disease. Asbestosis is observed in approximately 200,000 patients annually and asbestos-related deaths are estimated at 4,000 annually1. Although advances have been made using single gene/gene product or pathway studies, the complexity of the response to asbestos and the many unanswered questions suggested the need for a systems biology approach. The objective of this study was to generate a comprehensive view of the transcriptional changes induced by crocidolite asbestos in A549 human lung epithelial cells. Results A statistically robust, comprehensive data set documenting the crocidolite-induced changes in the A549 transcriptome was collected. A systems biology approach involving global observations from gene ontological analyses coupled with functional network analyses was used to explore the effects of crocidolite in the context of known molecular interactions. The analyses uniquely document a transcriptome with function-based networks in cell death, cancer, cell cycle, cellular growth, proliferation, and gene expression. These functional modules show signs of a complex interplay between signaling pathways consisting of both novel and previously described asbestos-related genes/gene products. These networks allowed for the identification of novel, putative crocidolite-related genes, leading to several new hypotheses regarding genes that are important for the asbestos response. The global analysis revealed a transcriptome that bears signatures of both apoptosis/cell death and cell survival/proliferation. Conclusion Our analyses demonstrate the power of combining a statistically robust, comprehensive dataset and a functional network genomics approach to 1 identify and explore relationships between genes of known importance

  7. Sulindac enhances arsenic trioxide induced apoptotic potential mediated by reactive oxygen species production in arsenic trioxide-resistant A549 lung carcinoma cells

    Full text: Recent reports indicate a broad spectrum of antitumor activity for arsenic trioxide (As2 O3) due to its ability to induce apoptosis via intracellular production of reactive oxygen species (ROS). Sulindac and nonsteroidal anti-inflammatory drugs induce apoptosis in a variety of cancer cells, including those of colon, prostate, breast, and leukemia. Therefore, we examined the effects of sulindac on As2O3-induced apoptosis in As2 O3-resistant A549 lung carcinoma cells in clinically available concentrations. Sulindac produced hydrogen peroxide (H 2 O 2 ) and nitric oxide (NO) in a dose-dependent manner and greatly sensitized the cells to As2O3-induced apoptosis. Apoptotic cell death was preceded by collapse of the mitochondrial membrane potential, release of cytochrome c/apoptosis inducing factor(AIF) and activation of caspase-3, -8, -9 activation. Importantly, the combined effect of As2O3 and sulindac was associated with an increased production of intracellular H2O3/reactive nitrogen species(RNS) and was completely suppressed by the reduced glutathione. In conclusion, intracellular ROS/RNS products most likely constitute the key mediators contributing to the combined effect of As2O3 and sulindac. Our data provide evidence for the first time that sulindac may help to extend the therapeutic spectrum of As2O3 and suggest that the combination of As2O3 and sulindac could be more broadly applied in cancer therapy

  8. Lysosome-associated membrane glycoprotein 3 is involved in influenza A virus replication in human lung epithelial (A549 cells

    Wang Jianwei

    2011-08-01

    Full Text Available Abstract Background Influenza A virus mutates rapidly, rendering antiviral therapies and vaccines directed against virus-encoded targets ineffective. Knowledge of the host factors and molecular pathways exploited by influenza virus will provide further targets for novel antiviral strategies. However, the critical host factors involved in influenza virus infection have not been fully defined. Results We demonstrated that LAMP3, a member of lysosome-associated membrane glycoprotein (LAMP family, was significantly induced in human lung epithelial (A549 cells upon influenza A virus infection. Knockdown of LAMP3 expression by RNA interference attenuated production of viral nucleoprotein (NP as well as virus titers. Confocal microscopy results demonstrated that viral NP is colocalized within LAMP3 positive vesicles at early stages of virus infection. Furthermore, knockdown of LAMP3 expression led to a reduction in nuclear accumulation of viral NP and impeded virus replication. Conclusions LAMP3 is an influenza A virus inducible gene, and plays an important role in viral post-entry steps. Our observations may provide insights into the mechanism of influenza virus replication and potential targets for novel anti-influenza therapeutics.

  9. Mitochondrial transcription factor A regulated ionizing radiation-induced mitochondrial biogenesis in human lung adenocarcinoma A549 cells

    Mitochondrial transcription factor A (TFAM), the first well-characterized transcription factor from vertebrate mitochondria, is closely related to mitochondrial DNA (mtDNA) maintenance and repair. Recent evidence has shown that the ratio of mtDNA to nuclearDNA (nDNA) is increased in both human cells and murine tissues after ionizing radiation (IR). However, the underlying mechanism has not as yet been clearly identified. In the present study, we demonstrated that in human lung adenocarcinoma A549 cells, expression of TFAM was upregulated, together with the increase of the relative mtDNA copy number and cytochrome c oxidase (COX) activity after α-particle irradiation. Furthermore, short hairpin RNA (shRNA)-mediated TFAM knockdown inhibited the enhancement of the relative mtDNA copy number and COX activity caused by α-particles. Taken together, our data suggested that TFAM plays a crucial role in regulating mtDNA amplification and mitochondrial biogenesis under IR conditions. (author)

  10. 紫杉醇奥曲肽偶合物对A549裸鼠移植瘤的靶向治疗研究%Tumor growth Inhibition of paclitaxel-octreotide conjugates on human non small cell lung cancer: experiment with mice

    申洪昌; 王秀问; 刘延国; 王亚伟; 魏军民; 马道新; 王朴; 李蕾; 孙美丽

    2008-01-01

    Objective To evaluate the antitumor effects of the conjugates synthesized by coupling cytotoxlc paclitaxel (PTX) to somatostatin analog octreotide (OCT) on human non small cell lung cancer (NSCLC). Methods Two cytotoxic somatostatin analog, PTX-OCT and 2PTX-OCT, were developed in which PIX was linked to octreotide. Forty-five BALB/c nu/nu nude mice were injected with human NSCLC cells of the line A549 into the fight armpit The nude mice that were xenografted were randomly divided into 8 groups. ①control group (n=6), ②FIX-OCT group (n=5), injected intravenously with PTC-OCT 150 nmol/kg on days 1,7, and 21, ③ 2PTX-OCT group (n=6), injected intravenously with PTrc-ocT 150 nmoL/kg, ④ OP group (n=6), injected with mixture of FIX and OCT 150 nmol/kg, ⑤ OCT group (n=5) injected with OCT 150 nmoL/kg ⑤ PTX group (n=6), injected with PTX 150 nmoL/kg, ⑦ 2PTX group, injected with PTX 300 nmol/kg, and ⑧2(PTX-OCT) injected with PTX-OCT 300 nmol/kg, The tumor volume and body weight (BW) were observed regularly. The tumor volume doubling time was calculated. White blood cells were counted by collecting peripheral blood sample. By the end of experiment the mice were killed with the tumors taken out. The expression of mRNA of subtypesl-5 of human somatostatin receptor (SSTR1-SSTR5 ) were investigated using RT-PCR Histological apoptosis was detected by DNA ladder. Immunohistoehemistry was used to examine the SSTR2 and SSTR5 expression and tumor microvessel density (MVD). Results The tumor volumes of 2PTX-OCT and 2 (2PTX-OCT) groups were significantly smaller than those of other groups (all P< 0.01 ). The tumor doubling times of the 2tPTX-OCT and 2(2PTX-OCT) groups were significantly longer than those of the other groups too (al.1P<0.01). The MVD levels of the 2tTX-OCT and 2(2PTX-OCT) groups were significant lower than those of the other groups (all P<0.01 ). The toxicity of the PTX group was more obvious. The WBC count levels of the PTX and 2PTX groups were

  11. Hypoxia in models of lung cancer

    Graves, Edward E; Vilalta, Marta; Cecic, Ivana K;

    2010-01-01

    PURPOSE: To efficiently translate experimental methods from bench to bedside, it is imperative that laboratory models of cancer mimic human disease as closely as possible. In this study, we sought to compare patterns of hypoxia in several standard and emerging mouse models of lung cancer to...... establish the appropriateness of each for evaluating the role of oxygen in lung cancer progression and therapeutic response. EXPERIMENTAL DESIGN: Subcutaneous and orthotopic human A549 lung carcinomas growing in nude mice as well as spontaneous K-ras or Myc-induced lung tumors grown in situ or......H2AX foci in vitro and in vivo. Finally, our findings were compared with oxygen electrode measurements of human lung cancers. RESULTS: Minimal fluoroazomycin arabinoside and pimonidazole accumulation was seen in tumors growing within the lungs, whereas subcutaneous tumors showed substantial trapping...

  12. Effects of Alpha Particle and Proton Beam Irradiation as Putative Cross-Talk between A549 Cancer Cells and the Endothelial Cells in a Co-Culture System

    Riquier, Hélène; Abel, Denis [URBC-NARILIS, University of Namur, 61 rue de Bruxelles, Namur 5000 (Belgium); Wera, Anne-Catherine; Heuskin, Anne-Catherine [LARN-PMR, NARILIS, University of Namur, Namur 5000 (Belgium); Genard, Géraldine [URBC-NARILIS, University of Namur, 61 rue de Bruxelles, Namur 5000 (Belgium); Lucas, Stéphane [LARN-PMR, NARILIS, University of Namur, Namur 5000 (Belgium); Michiels, Carine, E-mail: carine.michiels@unamur.be [URBC-NARILIS, University of Namur, 61 rue de Bruxelles, Namur 5000 (Belgium)

    2015-03-18

    Background: High-LET ion irradiation is being more and more often used to control tumors in patients. Given that tumors are now considered as complex organs composed of multiple cell types that can influence radiosensitivity, we investigated the effects of proton and alpha particle irradiation on the possible radioprotective cross-talk between cancer and endothelial cells. Materials and Methods: We designed new irradiation chambers that allow co-culture study of cells irradiated with a particle beam. A549 lung carcinoma cells and endothelial cells (EC) were exposed to 1.5 Gy of proton beam or 1 and 2 Gy of alpha particles. Cell responses were studied by clonogenic assays and cell cycle was analyzed by flow cytometry. Gene expression studies were performed using Taqman low density array and by RT-qPCR. Results: A549 cells and EC displayed similar survival fraction and they had similar cell cycle distribution when irradiated alone or in co-culture. Both types of irradiation induced the overexpression of genes involved in cell growth, inflammation and angiogenesis. Conclusions: We set up new irradiation chamber in which two cell types were irradiated together with a particle beam. We could not show that tumor cells and endothelial cells were able to protect each other from particle irradiation. Gene expression changes were observed after particle irradiation that could suggest a possible radioprotective inter-cellular communication between the two cell types but further investigations are needed to confirm these results.

  13. Effects of Alpha Particle and Proton Beam Irradiation as Putative Cross-Talk between A549 Cancer Cells and the Endothelial Cells in a Co-Culture System

    Background: High-LET ion irradiation is being more and more often used to control tumors in patients. Given that tumors are now considered as complex organs composed of multiple cell types that can influence radiosensitivity, we investigated the effects of proton and alpha particle irradiation on the possible radioprotective cross-talk between cancer and endothelial cells. Materials and Methods: We designed new irradiation chambers that allow co-culture study of cells irradiated with a particle beam. A549 lung carcinoma cells and endothelial cells (EC) were exposed to 1.5 Gy of proton beam or 1 and 2 Gy of alpha particles. Cell responses were studied by clonogenic assays and cell cycle was analyzed by flow cytometry. Gene expression studies were performed using Taqman low density array and by RT-qPCR. Results: A549 cells and EC displayed similar survival fraction and they had similar cell cycle distribution when irradiated alone or in co-culture. Both types of irradiation induced the overexpression of genes involved in cell growth, inflammation and angiogenesis. Conclusions: We set up new irradiation chamber in which two cell types were irradiated together with a particle beam. We could not show that tumor cells and endothelial cells were able to protect each other from particle irradiation. Gene expression changes were observed after particle irradiation that could suggest a possible radioprotective inter-cellular communication between the two cell types but further investigations are needed to confirm these results

  14. Effects of Alpha Particle and Proton Beam Irradiation as Putative Cross-Talk between A549 Cancer Cells and the Endothelial Cells in a Co-Culture System

    Hélène Riquier

    2015-03-01

    Full Text Available Background: High-LET ion irradiation is being more and more often used to control tumors in patients. Given that tumors are now considered as complex organs composed of multiple cell types that can influence radiosensitivity, we investigated the effects of proton and alpha particle irradiation on the possible radioprotective cross-talk between cancer and endothelial cells. Materials and Methods: We designed new irradiation chambers that allow co-culture study of cells irradiated with a particle beam. A549 lung carcinoma cells and endothelial cells (EC were exposed to 1.5 Gy of proton beam or 1 and 2 Gy of alpha particles. Cell responses were studied by clonogenic assays and cell cycle was analyzed by flow cytometry. Gene expression studies were performed using Taqman low density array and by RT-qPCR. Results: A549 cells and EC displayed similar survival fraction and they had similar cell cycle distribution when irradiated alone or in co-culture. Both types of irradiation induced the overexpression of genes involved in cell growth, inflammation and angiogenesis. Conclusions: We set up new irradiation chamber in which two cell types were irradiated together with a particle beam. We could not show that tumor cells and endothelial cells were able to protect each other from particle irradiation. Gene expression changes were observed after particle irradiation that could suggest a possible radioprotective inter-cellular communication between the two cell types but further investigations are needed to confirm these results.

  15. Curcumin Promoted the Apoptosis of Cisplain-resistant Human Lung Carcinoma Cells A549/DDP through Down-regulating miR-186*

    Jian ZHANG

    2010-04-01

    Full Text Available Background and objective Curcumin, a natural compound, is derived from the rthizom of Curcuma longa. In vitro and in vivo preclinical studies have shown its anti-inflammatory, antioxidant, anticancer activities and so on. miR-186*, which was found by microarray technology, was highly expressed in lung carcinoma cells A549/DDP. The aim of this study is to illustrate whether Curcumin could promote the apoptosis of A549/DDP cells through regulating the expression of miR-186*. Methods An oligonucleotide microarray chip was used to profile microRNA (miRNA expressions in A549/DDP cells treated with and without Curcumin. The significantly differentially expressed miRNA, which was selected from microarray chip, validated by quantitative real-time PCR. Ultimately, the remarkably expressed miRNA modulated the apoptosis assaying by flow cytometry expriments and the survival rate was measured by MTT method. Results The microarray chip results demonstrated: Curcumin altered the expression level of miRNAs compared with untreated control in A549/DDP cell line, miR-186* was significantly down-regulated after Curcumin treatment, which confirmed by quantitative real-time PCR. Downregulation of miR-186* expression by curcumin elevated the apoptosis, and the survival rate of A549/DDP cells decreased; but up-regulation of miR-186* expression by transfection its mimics restrained the apoptosis, the survival rate of A549/DDP cells increased, which were assayed by flow cytometry expriments and MTT method. Conclusion Modulation of miRNAs expression may be an important mechanism underlying the biological roles of Curcumin.

  16. Crocus sativus L. (Saffron) Stigma Aqueous Extract Induces Apoptosis in Alveolar Human Lung Cancer Cells through Caspase-Dependent Pathways Activation

    Saeed Samarghandian; Abasalt Borji; Seyed Kazem Farahmand; Reza Afshari; Saeideh Davoodi

    2013-01-01

    Worldwide, lung cancer is the most common form of cancer. Saffron has been used in folk medicine for centuries. We investigated the potential of saffron to induce cytotoxic and apoptotic effects in lung cancer cells (A549). We also examined the caspase-dependent pathways activation of saffron-induced apoptosis against the A549 cells. A549 cells were incubated with different concentrations of saffron extract; then cell morphological changes, cell viability, and apoptosis were determined by the...

  17. Polyurethane dispersion containing quaternized ammonium groups: An efficient nanosize gene delivery carrier for A549 cancer cell line transfection.

    Yousefpour Marzbali, Mahsa; Yari Khosroushahi, Ahmad; Movassaghpour, AliAkbar; Yeganeh, Hamid

    2016-01-25

    A novel polyurethane containing cationic ammonium groups (QPU) was synthesized and used as vector for gene therapy and cancer gene targeting. The synthesized QPU was characterized by Fourier transform infrared and nuclear magnetic resonance spectroscopy methods. An agarose gel retardation electrophoresis assay was conducted to verify the complete complex formation between QPU and pDNA. The particles size and zeta potential of neat polymers, plasmid DNA, polymers/DNA polyplexes were determined by the dynamic light scattering technique. The polyplexes cytotoxicity was determined using [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and its transfection efficiency was examined qualitatively by fluorescent microscopy and quantitatively by flow cytometery methods. The gel retardation assay, particle size and zeta potential measurements were confirmed that the synthesized cationic polymer could condense DNA efficiently in the physiologic condition. QPU polyplexes showed a significantly lower cytotoxicity compared to Polyfect polyplexes in the examined human cancerous (A549) or normal cells (KDR). Based on our findings, the transfection efficiency by QPU was 2.2 fold higher than Polyfect in the A549 cells whereas in the KDR cells, the cell transfection by Polyfect was 18.1 fold higher than QPU. Due to low cytotoxicity for normal cells and high transfection efficiency in cancer cells, the potential applicability of designed QPU as a non-viral gene carrier for targeting of cancer gene therapy was confirmed. PMID:26658031

  18. Exosome cargo reflects TGF-β1-mediated epithelial-to-mesenchymal transition (EMT) status in A549 human lung adenocarcinoma cells.

    Kim, Jiyeon; Kim, Tae Yeon; Lee, Myung Shin; Mun, Ji Young; Ihm, Chunhwa; Kim, Soon Ae

    2016-09-16

    It has been suggested that tumor cells secrete exosomes to modify the local microenvironment, which then promotes intercellular communication and metastasis. Although exosomes derived from cancer cells may contribute to the epithelial-mesenchymal transition (EMT) in untransformed cells, few studies have defined exosome cargo upon induction of EMT. In this study, we investigated the changes in exosomal cargo from the epithelial to mesenchymal cell phenotype by inducing EMT with transforming growth factor (TGF)-β1 in A549 human lung adenocarcinoma cells. The protein content of the exosomes reflects the change in the cell phenotype. In addition, miR-23a was significantly enriched in the exosomes after mesenchymal transition. Following treatment of exosomes from mesenchymal cells via EMT induction with TGF-β1 to the epithelial cell type, phenotypic changes in protein expression level and cell morphology were observed. Autologous treatment of exosomes enhanced the transcriptional activity and abundance of β-catenin. Our results suggest that the exosomal protein and miRNA content reflects the physiological condition of its source and that exosomes induce phenotypic changes via autocrine signaling. PMID:27492069

  19. Effect of functionalized and non-functionalized nanodiamond on the morphology and activities of antioxidant enzymes of lung epithelial cells (A549).

    Solarska-Ściuk, Katarzyna; Gajewska, Agnieszka; Glińska, Sława; Michlewska, Sylwia; Balcerzak, Łucja; Jamrozik, Agnieszka; Skolimowski, Janusz; Burda, Květoslava; Bartosz, Grzegorz

    2014-10-01

    The development of nanotechnology opens up new ways for biomedical applications of unmodified and modified diamond nanoparticles which are one of the most popular nanomaterials used in biology, biotechnology, medicine, cosmetics and engineering. They have been applied as diagnostic and therapeutic agents because they can be targeted to and localized in cells causing apoptosis and necrosis. The problem of biocompatibility of nanodiamonds at higher concentrations is thus of primary importance. The first step in the modification of DNPs is usually the introduction of hydrogen groups, which can bind other functional groups. The basic method to introduce -OH groups onto nanoparticles is the Fenton reaction. The aim of this study was to compare the effect of unmodified nanodiamond particles and nanoparticles modified by introduction of -OH groups and etoposide onto their surface reaction on human non-small lung cancer cells. A549 cells were incubated with 2-100μg/ml nanopowders and at 0.6-24μg/ml etoposide in the DMEM medium. We observed a decrease of cells viability and generation of reactive oxygen/ nitrogen species in the cells after incubation, estimated by oxidation of H2DCF-DA and DAF-FM-DA. Modified detonation nanoparticles affected also the cellular content of glutathione and activities of main antioxidant enzymes (glutathione peroxidase, glutathione reductase, glutathione S-transferase, superoxide dismutase and catalase). The results of TEM microscopy show changes in cell morphology. These data demonstrate that modified nanoparticles induce oxidative stress in the target cells. PMID:25451571

  20. Treatment of transplanted tumor of lung adenocarcinoma A549 transfected by human somatostatin receptor subtype 2 (hsstr2) gene with 188Re-RC-160

    Background and aim: Radionuclide-labeled somatostatin analogues selectively target somatostatin receptor (SSTR)-expressing tumors as a basis for diagnosis and treatment of these tumors. To those tumors without somatostatin receptor expressed, the hSSTR2 gene was transfected. Express of the hSSTR2 receptor was imaging and the radiotherapeutic effect was evaluated with 188Re-RC-160. Methods: The stable hSSTR2-expressing A549 cells (pcDNA3-hSSTR2 A549) and non-somatostatin receptor expressing A549 cells (pcDNA3 A549) were selected by western blot. Later, a corresponding animal tumor model was established. Expression of the hSSTR2 reporter was imaged using 188Re-RC-160 recognition. Tumors were evaluated for somatostatin receptor expression using immunohistochemistry. The distribution of 188Re-RC-160 in the animal tumor model was measured and the inhibitory effects of 188Re-RC-160 were evaluated by measurement of tumor growth and hematoxylin and eosin and TdT mediated dUTP nick end labeling (TUNEL) staining. Results: In vivo radioimaging revealed specific targeting of 188Re-RC-160 to tumors derived from pcDNA3- hSSTR2 A549 cells, compared to those from pcDNA3 A549 cells. pcDNA3- hSSTR2 A549 tumor growth inhibition was significantly higher in the single 7.4 MBq 188Re-RC-160 treatment group than in the 2x7.4 MBq rhenium-188, RC-160 group, control group, and pcDNA3 A549 tumors (P188Re-RC-160), induced significantly increased tumor-growth inhibition compare with single 7.4 MBq 188Re-RC-160 treatment (P188Re-RC-160 could be effectively used for targeting therapy the A549-derived tumors exogenously expressing hSSTR2, which will offers a potential therapeutic strategy for the treatment of somatostatin receptor-negative cancers.

  1. Inhibitory effect of recombinant vector pEgr1-hsTRAIL induced by radiation on growth of lung adenocarcinoma A549 cells

    Objective: to construct human secreted TRAIL (hsTRAIL) recombinant vector pEgr1-hsTRAIL mediated by Egr1, and to explore the inhibitory effect on the growth of lung adenocarcinoma A549 cells. Methods: The hsTRAIL vet or mediated by Egr1 was constructed by gene recombination technique, the A549 cells were transfected with the plasmid after identification by PCR, restrictive enzyme digestion and sequencing, and irradiated by 6 Gy X-rays. There were control group, pEgr1-hsTRAIL group, 6 Gy X-rays group and pEgr1-hsTRAIL + 6 Gy X-rays group in the experiment. The expression of hsTRAIL in A549 cells was detected by ELISA method, the cell proliferation was detected by MTT assay, the cycle changes of cell cycle the detected by flow cytometry and the apoptosis was measured by TUNEL method. Results: The hsTRAIL recombinant vector pEgr1-hsTRAIL mediated by Egr1 was constructed successfully. The cells were irradiated by 6 Gy X-rays after transfected with plasmid. The hsTRAIL protein expressions in control, 6 Gy and pEgr1-hsTRAIL groups didn't change significantly with the time prolongation, but the expression in pEgr1-hsTRAIL + 6 Gy group was increased significantly with the time prolongation (P<0.05 or P<0.01), and reached to peak value at 8 h. There was no significant difference of A549 cell proliferation ability between control group and pEgr1-hsTRAIL group, but the proliferation abilities in 6 Gy and pEgr1-hsTRAIL + 6 Gy groups were decreased significantly compared with control group, especially in pEgr1-hsTRAIL + 6 Gy group (P<0.05 or P<0.01). Compared with control group, the percentages of A549 cells at different phases in pEgr1-hsTRAIL group didn't change significantly, but the percentages of A549 cells at G0/G1 phase in 6 Gy and pEgr1-hsTRAIL + 6 Gy groups were increased significantly (P<0.05), the percentages of A549 cells at G2/M phase were decreased significantly (P<0.05), the percentages of A549 cells at S phase didn't change significantly. The

  2. Rapid Cancer Fluorescence Imaging Using A γ-Glutamyltranspeptidase-Specific Probe For Primary Lung Cancer

    Hino, Haruaki; Kamiya, Mako; Kitano, Kentaro; Mizuno, Kazue; Tanaka, Sayaka; Nishiyama, Nobuhiro; Kataoka, Kazunori; Urano, Yasuteru; Nakajima, Jun

    2016-01-01

    BACKGROUND: We set out to examine the activity of γ-glutamyltranspeptidase (GGT) in lung cancer and the validity of γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) for intraoperative imaging of primary lung cancer. METHODS: GGT activities and mRNA expression levels of GGT1 (one of the GGT subtypes) in five human lung cancer cell lines were examined by fluorescence imaging and quantitative reverse transcription polymerase chain reaction. In vivo imaging of an orthotopic A549 xenograft mod...

  3. Effect of primarily cultured human lung cancer-associated fibroblasts on radiosensitivity of lung cancer cells

    Objective: To investigate the effect of human lung cancer-associated fibroblasts (CAF) on the radiosensitivity of lung cancer cells when CAF is placed in direct contact co-culture with lung cancer cells. Methods: Human lung CAF was obtained from fresh human lung adenocarcinoma tissue specimens by primary culture and subculture and was then identified by immunofluorescence staining. The CAF was placed in direct contact co-culture with lung cancer A549 and H1299 cells, and the effects of CAF on the radiosensitivity of A549 and H1299 cells were evaluated by colony-forming assay. Results: The human lung CAF obtained by adherent culture could stably grow and proliferate, and it had specific expression of α-smooth muscle actin, vimentin, and fibroblast activation protein,but without expression of cytokeratin-18. The plating efficiency (PE, %) of A549 cells at 0 Gy irradiation was (20.0 ± 3.9)% when cultured alone versus (32.3 ± 5.5)% when co-cultured with CAF (t=3.16, P<0.05), and the PE of H1299 cells at 0 Gy irradiation was (20.6 ± 3.1)% when cultured alone versus (35.2 ± 2.3)% when co-cultured with CAF (t=6.55, P<0.05). The cell survival rate at 2 Gy irradiation (SF2) of A549 cells was 0.727 ±0.061 when cultured alone versus 0.782 ± 0.089 when co-cultured with CAF (t=0.88, P>0.05), and the SF2 of H1299 cells was 0.692 ±0.065 when cultured alone versus 0.782 ± 0.037 when co-cultured with CAF (t=2.08, P>0.05). The protection enhancement ratios of human lung CAF for A549 cells and H1299 cells were 1.29 and 1.25, respectively. Conclusions: Human lung CAF reduces the radiosensitivity of lung cancer cells when placed in direct contact co-culture with them, and the radioprotective effect may be attributed to CAF promoting the proliferation of lung cancer cells. (authors)

  4. Effects of Hypoxia on Expression of P-gp and Mutltidrug Resistance Protein in Human Lung Adenocarcinoma A549 Cell Line

    XIA Shu; YU Shiying; YUAN Xianglin

    2005-01-01

    Summary: To study the effects of hypoxia on the expression of P-gp and mutltidrug resistance protein in human lung adenocarcinoma A549 cell line, and to explore the probable mechanism of hypoxia in tumor cell of MDR. The expression of hypoxia inducible factor-1α, P-gp and mutltidrug resistance protein was immunohistochemically detected by culturing human lung adenocarcinoma A549 cell under hypoxia (2 % O2) for 24 h. After interaction with adriamycin or cisplatin under hypoxia (2 % O2) for 24 h, the cell survival rate was detected by MTT. Our results showed that the expression of hypoxia inducible factor-1α, P-gp and mutltidrug resistance protein under hypoxia were higher than the expression under normoxia, and correlations between the expression of HIF-1α and P-gp or multidrug resistance-associated protein was observed (P<0.05). The resistance of adriamycin of A549 cell was enhanced under hypoxia. It is concluded that the resistance of tumor chemotherapy is enhanced in hypoxia. The expression of HIF-1α is obviously correlated with the expression of P-gp and mutltidrug resistance protein.

  5. Staging of Lung Cancer

    ... which therapy (or therapies) should be used. Second, lung cancer staging tells how much your cancer has spread. Knowing ... your body. How good are these tests at staging lung cancer? If your biopsy finds cancer cells, this is ...

  6. Okadaic acid inhibits cell multiplication and induces apoptosis in a549 cells, a human lung adenocarcinoma cell line

    Wang, Renjun; Lv, Lili; Zhao, Yunfeng; Yang, Nana

    2014-01-01

    This essay aims to research the effect of okadaic acid (OA) on A549 cell multiplication, and cell apoptosis induced by OA was observed by cell morphology. MTT assay, trypan blue exclusion test (TBET), Giemsa staining method and acridine orange (AO) fluorescence staining assay were applied. The results of cell survival evaluated by TBET and colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) showed: The number of A549 cells was decreased in a dose-depende...

  7. Establishment of a radioresistant human lung cancer cell subline and its mechanism of radioresistance

    Objective: To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods: Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays: A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones, together with its parental A549 cells were measured by clone formation assay and flow cytometry. The mRNA and protein levels of Notchl in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results: Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G1/G1 phase (P0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05). The expression of Notchl had no marked change compared to A549 cell. Conclusions: The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlaiton with the expression of Notchl. (authors)

  8. Chemoradiotherapy for lung cancer

    Lung cancer remains a disease associated with a poor prognosis. Chemoradiotherapy is performed for unresectable stage 3 non-small cell lung cancer (NSCLC) and inoperable limited-disease small cell lung cancer. In this lecture, chemoradiotherapy for lung cancer is outlined primary according to the 2014 edition of the Clinical Practice Guidelines for Lung Cancer, and also referring to molecular targeted drugs, radiation pneumonitis, and particle radiotherapy. (author)

  9. Low-dose carbon-based nanoparticle-induced effects in A549 lung cells determined by biospectroscopy are associated with increases in genomic methylation

    Li, Junyi; Tian, Meiping; Cui, Li; Dwyer, John; Fullwood, Nigel J.; Shen, Heqing; Martin, Francis L.

    2016-02-01

    Nanotechnology has introduced many manufactured carbon-based nanoparticles (CNPs) into our environment, generating a debate into their risks and benefits. Numerous nanotoxicology investigations have been carried, and nanoparticle-induced toxic effects have been reported. However, there remain gaps in our knowledge, primarily regarding mechanism. Herein, we assessed the global alterations induced by CNPs in A549 lung cells using biospectroscopy techniques, including attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy and surface-enhanced Raman spectroscopy (SERS). A549 cells were treated with fullerene (C60), long or short multi-walled carbon nanotubes, or single-walled carbon nanotubes at concentrations of 0.1 mg/L, 0.01 mg/L and 0.001 mg/L. Exposed cells were then analysed by ATR-FTIR spectroscopy and SERS. Spectra were pre-processed via computational analysis, and information on biochemical alterations in exposed cells were identified. Additionally, global DNA methylation levels in cells exposed to CNPs at 0.1 mg/L were determined using HPLC-MS and genetic regulators (for DNA methylation) were checked by quantitative real-time RT-PCR. It was found that CNPs exert marked effects in A549 cells and also contribute to increases in global DNA methylation. For the first time, this study highlights that real-world levels of nanoparticles can alter the methylome of exposed cells; this could have enormous implications for their regulatory assessment.

  10. Purification and characterization of protease enzyme from actinomycetes and its cytotoxic effect on cancer cell line (A549)

    C Balachandran; V Duraipandiyan; S Ignacimuthu

    2012-01-01

    Objective: To isolate active actinomycetes from soil samples of Northern Himalayas and study their culture characterization, protease production and cytotoxic effects on cancer cell line (A549). Methods: Forty six strains of actinomycetes were isolated from the soil collected from Northern Himalayas, India. Isolation of actinomycetes was performed by serial dilution plate technique. Forty six isolated actinomycetes cultures were grown in ISP 2 medium to study the morphology and biochemical characteristics. Isolated strains were studied for protease enzyme production in skim milk agar medium with solubilising capacity. Seven isolates were studied for melanin pigmentation and different NaCl concentration. Effects of environmental conditions influencing protease enzyme production of seven isolated strains were also studied at different pH, temperature and metal ions (β-mercaptoethanol, dithiothreitol, iodoacetamide, MgSO4, CaCl2 and EDTA). The seven isolates were also studied for lytic enzyme activity using different bacteria and yeast such as Pseudomonas aeruginosa (P. aeruginosa), Enterococcus feacalis (E. feacalis), Escherishia coli (E. coli), Candida albicans (C. albicans), Bacillus subtilis (B. subtilis), Klebsiella pneumonia (K. pneumonia) and Staphylococcus aureus (S. aureus). Results: Isolates ERIA-31 and ERIA-33 produced more protease enzyme activity in modified nutrient agar media compared to other actinomycetes cultures. ERIA-31 and ERIA-33 were tested for cytotoxic effect in human adenocarcinoma cancer cell line (A549). IC50 for ERIA-31 was 57.04 μg/mL and IC50 for ERIA-33 was 55.07 μg/mL. Conclusion: Actinomycete being a protease producing bacteria has the potential for use in industrial purpose, pharmaceuticals, cytotoxic agent and its proteolytic activity. Isolates of ERIA-31 and ERIA-33 produced significant amount of protease enzymes.

  11. Damaging and protective bystander cross-talk between human lung cancer and normal cells after proton microbeam irradiation

    Desai, Sejal [Radiation Signalling and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Kobayashi, Alisa; Konishi, Teruaki; Oikawa, Masakazu [Radiation System and Engineering Section, Department of Technical Support and Development, Research, Development and Support Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Pandey, Badri N., E-mail: badrinarain@yahoo.co.in [Radiation Signalling and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India)

    2014-05-15

    Graphical abstract: - Highlights: • Proton-microbeam irradiated A549 cells send damaging signals to bystander A549 cells. • Irradiated A549A549 bystander response is through gap junctional communication. • Bystander WI38 cells exert protective signalling in irradiated A549 cells. • Rescue of irradiated A549 cells by WI38 cells is independent of gap junctions. - Abstract: Most of the studies of radiation-induced bystander effects (RIBE) have been focused on understanding the radiobiological changes observed in bystander cells in response to the signals from irradiated cells in a normal cell population with implications to radiation risk assessment. However, reports on RIBE with relevance to cancer radiotherapy especially investigating the bidirectional and criss-cross bystander communications between cancer and normal cells are limited. Hence, in present study employing co-culture approach, we have investigated the bystander cross-talk between lung cancer (A549) and normal (WI38) cells after proton-microbeam irradiation using γ-H2AX foci fluorescence as a measure of DNA double-strand breaks (DSBs). We observed that in A549A549 co-cultures, irradiated A549 cells exert damaging effects in bystander A549 cells, which were found to be mediated through gap junctional intercellular communication (GJIC). However, in A549–WI38 co-cultures, irradiated A549 did not affect bystander WI38 cells. Rather, bystander WI38 cells induced inverse protective signalling (rescue effect) in irradiated A549 cells, which was independent of GJIC. On the other hand, in response to irradiated WI38 cells neither of the bystander cells (A549 or WI38) showed significant increase in γ-H2AX foci. The observed bystander signalling between tumour and normal cells may have potential implications in therapeutic outcome of cancer radiotherapy.

  12. Damaging and protective bystander cross-talk between human lung cancer and normal cells after proton microbeam irradiation

    Graphical abstract: - Highlights: • Proton-microbeam irradiated A549 cells send damaging signals to bystander A549 cells. • Irradiated A549A549 bystander response is through gap junctional communication. • Bystander WI38 cells exert protective signalling in irradiated A549 cells. • Rescue of irradiated A549 cells by WI38 cells is independent of gap junctions. - Abstract: Most of the studies of radiation-induced bystander effects (RIBE) have been focused on understanding the radiobiological changes observed in bystander cells in response to the signals from irradiated cells in a normal cell population with implications to radiation risk assessment. However, reports on RIBE with relevance to cancer radiotherapy especially investigating the bidirectional and criss-cross bystander communications between cancer and normal cells are limited. Hence, in present study employing co-culture approach, we have investigated the bystander cross-talk between lung cancer (A549) and normal (WI38) cells after proton-microbeam irradiation using γ-H2AX foci fluorescence as a measure of DNA double-strand breaks (DSBs). We observed that in A549A549 co-cultures, irradiated A549 cells exert damaging effects in bystander A549 cells, which were found to be mediated through gap junctional intercellular communication (GJIC). However, in A549–WI38 co-cultures, irradiated A549 did not affect bystander WI38 cells. Rather, bystander WI38 cells induced inverse protective signalling (rescue effect) in irradiated A549 cells, which was independent of GJIC. On the other hand, in response to irradiated WI38 cells neither of the bystander cells (A549 or WI38) showed significant increase in γ-H2AX foci. The observed bystander signalling between tumour and normal cells may have potential implications in therapeutic outcome of cancer radiotherapy

  13. Epidemiology of Lung Cancer

    Bahader Yasser; Jazieh Abdul-Rahman

    2013-01-01

    Incidence and mortality attributed to lung cancer has risen steadily since the 1930s. Efforts to improve outcomes have not only led to a greater understanding of the etiology of lung cancer, but also the histologic and molecular characteristics of individual lung tumors. This article describes this evolution by discussing the extent of the current lung cancer epidemic including contemporary incidence and mortality trends, the risk factors for development of lung cancer, and details of promisi...

  14. Radiation Therapy for Lung Cancer

    ... are available to help. HELPFUL WEB SITES ON LUNG CANCER American Lung Association www.lung.org Lungcancer.org www.lungcancer.org Lung Cancer Alliance www.lungcanceralliance.org Lung Cancer Online www. ...

  15. Epidemiology of Lung Cancer.

    Mao, Yousheng; Yang, Ding; He, Jie; Krasna, Mark J

    2016-07-01

    Lung cancer has been transformed from a rare disease into a global problem and public health issue. The etiologic factors of lung cancer become more complex along with industrialization, urbanization, and environmental pollution around the world. Currently, the control of lung cancer has attracted worldwide attention. Studies on the epidemiologic characteristics of lung cancer and its relative risk factors have played an important role in the tertiary prevention of lung cancer and in exploring new ways of diagnosis and treatment. This article reviews the current evolution of the epidemiology of lung cancer. PMID:27261907

  16. Influence of pEgr1-hsTRAIL plasmid on radiosensitivity and DR4 and DR5 expression levels in lung adencarcinoma A549 cells

    Objective: To measure the changes of the radiosensitivity in human lung adenocarcinoma A549 cells transfected with pEgr1-hsTRAIL plasmid and the effect on death receptor (DR) 4 and DR5 expressions, and to explore the radiosensitizing effect of pEgr1-hsTRAIL plasmid and possible mechanism on inducing apoptosis. Methods: There were normal control, pEgr1-hsTRAIL, 6 Gy X-rays, and pEgr1-hsTRAIL + 6 Gy X-rays groups in the experiment. After the A549 cells were transfected with liposome, and irradiated with X-rays, colony formation assay was used to measure the radiosensitivity, and reverse transcription PCR (RT-PCR) was performed to detect the DR4 and DR5 mRNA expressions, and Western blotting was applied to determine the DR4 and DR5 protein expressions. Results: The D0 values of A549 cells in normal control group and pEgr1-hsTRAIL group were 3.26 and 1.91 Gy, respectively, it indicated that pEgr1-hsTRAIL plasmid could enhance the radiosensitivity in A549 cells. The RT-PCR results showed that as compared with normal control group, the DR4 and DR5 mRNA expression levels in pEgr1-hsTRAIL group had no significant change, but those in 6 Gy X-rays group were increased significantly (P<0.05), and those in pEgr1-hsTRAIL + 6 Gy X-rays group were also increased significantly (P<0.05); the DR5 mRNA expression level in pEgr1-hsTRAIL + 6 Gy X-rays group was higher than that in 6 Gy X-rays group (P<0.05). The Western blotting results showed that the DR4 and DR5 protein expressions in pEgr1-hsTRAIL group did not change obviously compared with normal control group, but those in 6 Gy X-rays and pEgr1-hsTRAIL + 6 Gy X-rays groups were increased, and the DR5 protein expression in pEgr1-hsTRAIL + 6 Gy X-rays group was increased mostly. Conclusion: The recombinant plasmid pEgr1-hsTRAIL can enhance the radiosensitivity of A549 cells, and has the enhancing effect on DR5 expression induced by radiation, but no same effect on DR4 expression. (authors)

  17. Induction of COX-2 protein expression by vanadate in A549 human lung carcinoma cell line through EGF receptor and p38 MAPK-mediated pathway

    Vanadate is a transition metal widely distributed in the environment. It has been reported that vanadate associated with air pollution particles can modify DNA synthesis, causing cell growth arrest, and apoptosis. Moreover, vanadium exposure was also found to cause the synthesis of inflammatory cytokines, such as interleukin-1, tumor necrosis factor-α, and prostaglandin E2. Here, we found that exposure of A549 human lung carcinoma cells to vanadate led to extracellular signal-regulated kinase, c-Jun NH2-terminal protein kinases (JNKs), p38 mitogen-activated protein kinase (p38) activation, and COX-2 protein expression in a dose-dependent manner. SB203580, a p38 MAPK inhibitor, but not PD098059 and SP600125, specific inhibitor of MKK1 and selective inhibitor of JNK, respectively, suppressed COX-2 expression. Furthermore, the epithelial growth factor (EGF) receptor specific inhibitor (PD153035) reduced vanadate-induced COX-2 expression. However, scavenging of vanadate-induced reactive oxygen species by catalase, a specific H2O2 inhibitor, or DPI, an NADPH oxidase inhibitor, resulted in no inhibition on COX-2 expression. Together, we suggested that EGF receptor and p38 MAPK signaling pathway may be involved in vanadate-induced COX-2 protein expression in A549 human lung carcinoma cell line

  18. Lung and Bronchus Cancer

    ... copy to myself The information used on this page will not be used to send unsolicited emails or shared with a third party. HPF: Did You Know? Video Series Lung Cancer - Did you know that lung cancer is the ...

  19. Study on radiosensitivity by targeting HIF-1α in human lung cancer and growth of the transplanted tumors

    Objective: To observe the radiosensitivity by targeting HIF-lα in human lung cancer and the effects on tumor growth in nude mice. Methods: Radiosensitivity of A549 and A549/HIF-1α (-) cells were tested by clonogenic forming assay. A549/HIF-1 α (-) cells and A549 cells were injected into the male BALB/C nude mice. Tumor growth was observed. The expression of HIF-1α and microvessel density were detected by immunohistochemistry method. Results: SERs of HIF-1α gene silencing were 1.03 in normoxia and 1.65 in hypoxia.The sizes of tumor xenografts derived from A549/HIF-1α (-) cells were significantly reduced compared to those of the xenografts derived from A549 cells. HIF-1 α protein staining result showed a dramatic decrease in tumors from A549/HIF-1α (-) mice. The microvessel densities (MVD) were 19.83 ± 4.09 in A549 group and 11.61 ±3.04 in A549/HIF-lα (-) group (F=15.57, P<0.05). Conclusions: Hypoxia-induced radio-resistance in lung cancer A549 cells could be reversed by silencing the HIF-1α. It also retards the growth of tumor xenografts, decreases HIF-1α expression and reduces the vascularity. (authors)

  20. Enhancement of radiosensitivity by topoisomerase II inhibitor, amrubicin and amrubicinol, in human lung adenocarcinoma A549 cells and kinetics of apoptosis and necrosis induction.

    Hayashi, Sachiko; Hatashita, Masanori; Matsumoto, Hideki; Shioura, Hiroki; Kitai, Ryuhei; Kano, Eiichi

    2006-11-01

    The effects of amrubicin (AMR) and its active metabolite, amrubicinol (AMROH), on the sensitivity of human lung adenocarcinoma A549 cells to ionizing radiation were investigated in vitro. Further, the kinetics of apoptosis and necrosis induction were also analyzed. The cytocidal effects of X-ray irradiation on A549 cells resulted in a low level of radiosensitivity with a D0 value of 12 Gy. The slopes of the survival curves in the exponential phase were plotted on semilogarithmic paper for radiation combined with AMR (2.5 microg/ml) and AMROH (0.02 microg/ml) treatment, and were shown to be approximately parallel to treatment with irradiation alone. The initial shoulder-shape portion of the survival curve for radiation alone, indicating the repair of sublethal damage, was reduced as compared to that for sequential combined treatment with AMR or AMROH. Sequential treatments with AMR or AMROH prior to ionizing radiation resulted in an additive radio-enhancement effect that reduced not only survival, but also the shoulder width. Fractionated irradiation with 2 Gy per fraction of A549 cells was carried out in vitro similar to that commonly performed in clinical radiotherapy and the radio-resistance of the cells was shown to be inhibited by AMR and AMROH. Similar to AMR and AMROH, adriamycin and etoposide (VP-16) are DNA topoisomerase II inhibitors. The effects of these 4 agents on cells that received X-ray irradiation were compared and all of the agents exhibited comparable radio-enhancement effects. The induction of apoptosis was investigated at 48 and 72 h after administration of AMROH, radiation or combined treatment, and apoptosis was not significantly induced after any of the treatments. We also examined the induction of necrosis, and found that the incidence of necrosis following combined treatment was approximately 2 times higher than that with either of the single treatments. PMID:17016621

  1. CD147 deficiency blocks IL-8 secretion and inhibits lung cancer-induced osteoclastogenesis

    Bone is a frequent target of lung cancer metastasis, which is associated with significant morbidity and poor prognosis; however, the molecular basis of this process is still unknown. This study investigated the role of extracellular matrix metalloproteinase inducer (also known as cluster of differentiation (CD)147) in osteoclastogenesis resulting from bone metastasis, based on the enrichment of this glycoprotein on the surface of many malignant bone tumors. RNA interference was used to silence CD147 expression in A549 human lung cancer cells. Compared with conditioned medium (CM) from control cells (A549-CM), CM from CD147-deficient cells (A549-si-CM) suppressed receptor activator of nuclear factor κB ligand-stimulated osteoclastogenesis in RAW 264.7 cells and bone marrow-derived macrophages. The mRNA levels of osteoclast-specific genes such as tartrate-resistant acid phosphatase, calcitonin receptor, and cathepsin K were also reduced in the presence of A549-si-CM. CD147 knockdown in A549 cells decreased interleukin (IL)-8mRNA and protein expression. IL-8 is present in large amounts in A549-CM and mimicked its inductive effect on osteoclastogenesis; this was reversed by depletion of IL-8 from the medium. Taken together, these results indicate that CD147 promotes lung cancer-induced osteoclastogenesis by modulating IL-8 secretion, and suggest that CD147 is a potential therapeutic target for cancer-associated bone resorption in lung cancer patients. - Highlights: • Bone loss frequently results from lung cancer metastasis. • Cluster of differentiation (CD)147 was depleted in A549 lung adenocarcinoma cells. • RAW 264.7 cell osteoclastogenesis was blocked by medium from CD147-deficient cells. • Interleukin (IL)-8 level was reduced in the conditioned medium. • Osteoclastogenesis induced by lung tumor cells requires CD147-mediated IL-8 release

  2. CD147 deficiency blocks IL-8 secretion and inhibits lung cancer-induced osteoclastogenesis

    Wang, Hongkai; Zhuo, Yunyun; Hu, Xu; Shen, Weiwei; Zhang, Ying; Chu, Tongwei, E-mail: chtw@sina.com

    2015-03-06

    Bone is a frequent target of lung cancer metastasis, which is associated with significant morbidity and poor prognosis; however, the molecular basis of this process is still unknown. This study investigated the role of extracellular matrix metalloproteinase inducer (also known as cluster of differentiation (CD)147) in osteoclastogenesis resulting from bone metastasis, based on the enrichment of this glycoprotein on the surface of many malignant bone tumors. RNA interference was used to silence CD147 expression in A549 human lung cancer cells. Compared with conditioned medium (CM) from control cells (A549-CM), CM from CD147-deficient cells (A549-si-CM) suppressed receptor activator of nuclear factor κB ligand-stimulated osteoclastogenesis in RAW 264.7 cells and bone marrow-derived macrophages. The mRNA levels of osteoclast-specific genes such as tartrate-resistant acid phosphatase, calcitonin receptor, and cathepsin K were also reduced in the presence of A549-si-CM. CD147 knockdown in A549 cells decreased interleukin (IL)-8mRNA and protein expression. IL-8 is present in large amounts in A549-CM and mimicked its inductive effect on osteoclastogenesis; this was reversed by depletion of IL-8 from the medium. Taken together, these results indicate that CD147 promotes lung cancer-induced osteoclastogenesis by modulating IL-8 secretion, and suggest that CD147 is a potential therapeutic target for cancer-associated bone resorption in lung cancer patients. - Highlights: • Bone loss frequently results from lung cancer metastasis. • Cluster of differentiation (CD)147 was depleted in A549 lung adenocarcinoma cells. • RAW 264.7 cell osteoclastogenesis was blocked by medium from CD147-deficient cells. • Interleukin (IL)-8 level was reduced in the conditioned medium. • Osteoclastogenesis induced by lung tumor cells requires CD147-mediated IL-8 release.

  3. Experimental Study on A549 Cell Death Mediated by Xenoantigen α-gal 
in Human Serum

    Shengming ZHU

    2012-11-01

    Full Text Available Background and objective The absence of α-gal in humans is caused by the inactivity of α-1,3GT gene. However, humans have pre-existing and abundant anti-gal antibodies. Xenotransplantation procedures have indicated the high potential of introducing α-1,3GT gene to synthesize α-gal for cancer gene therapy by mimicking hyper-acute rejection. The aim of this study is to construct a lung cancer A549 cell line that expressed α-gal, and to observe the antitumor mechanisms mediated by human serum. Methods A549 cells were transfected with pEGFP-N1-GT plasmids constructed in a previous study. A stable transgenic cell line, A549-GT, was then selected and cultivated. The biological characteristics of A549-GT cells, including morphology and proliferation, were examined. α-1,3GT mRNA expression was detected by RT-PCR. Direct immunofluorescence staining and flow cytometry (FCM were used to analyze the synthesis of α-gal in A549-GT. The binding of human serum IgM and C3 with A549-GT were also detected. Results α-1,3GT mRNA was expressed in A549-GT. Direct immunofluorescence staining and FCM indicated a high and stable α-gal expression rate in A549-GT. Compared with parental A549 cells, the biological characteristics of A549-GT were unaltered. α-Gal expression was not detected in the human fetal lung fibroblast cell line MRC-5 even though A549-GT and its culture medium were cultivated with the enzyme. Immunofluorescence staining and FCM also indicated abundant binding between A549-GT treated with human serum and IgM/C3. Conclusion α-Gal expression in tumor cells by gene transduction can induce complement-dependent cytototic antitumor effects.

  4. Biological impacts of TiO2 on human lung cell lines A549 and H1299: particle size distribution effects

    Increasing use of titanium dioxide (TiO2) nanoparticles in many commercial applications has led to emerging concerns regarding the safety and environmental impact of these materials. In this study, we have investigated the biological impact of nano-TiO2 (with particle primary size of 20 nm Aeroxide P25) on human lung cell lines in vitro and also the effect of particle size distribution on the particle uptake and apparent toxicity. The biological impact of nano-TiO2 is shown to be influenced by the concentration and particle size distribution of the TiO2 and the impact was shown to differ between the two cell lines (A549 and H1299) investigated herein. A549 cell line was shown to be relatively resistant to the total amount of TiO2 particles uptaken, as measured by cell viability and metabolic assays, while H1299 had a much higher capacity to ingest TiO2 particles and aggregates, with consequent evidence of impact at concentrations as low as 30–150 μg/mL TiO2. Evidence gathered from this study suggests that both viability and metabolic assays (measuring metabolic and mitochondrial activities and also cellular ATP level) should be carried out collectively to gain a true assessment of the impact of exposure to TiO2 particles.

  5. Endostar combined with cryoablation for subcutaneous xenografted tumor model of lung adenocarcinoma cell line A549 in BALB/c nude mice: an experimental study

    Objective: To investigate the inhibitory effect of Endostar combined with cryoablation on Lung adenocarcinoma cell line A549 in BALB/c nude mice, and to discuss its interaction mechanisms. Methods: The lung adenocarcinoma A549 model in BALB/c nude mice were established. When the largest diameter of tumor reached 1.0 cm, a total of 24 mice were randomly and equally divided into 4 groups: control group, Endostar group, cryoablation group and cryoablation plus Endostar group. The largest diameter and the vertical diameter of the tumors were measured at different points of time after treatment. At the 21st day, the mice were sacrificed and the tumors were removed and the rate of tumor cell apoptosis, the microvessel density (MVD) and the expression level of vascular endothelial growth factor (VEGF) were determined by using immunohistochemistry method. The results were statistically analyzed. Results: The tumor growth velocity of the control group, Endostar group, cryoablation group and cryoablation plus Endostar group was (2.36.68±51.23)%, (220.02±30.61)%, (159.46±29.33)% and (103.34±25.50)%, respectively (P<0.01). The rate of apoptosis of the four groups was (21.67±2.34)%, (22.17±1.47)%, (38.33±1.37)% and (49.17±1.72)%, respectively (P<0.01). The MVD and the expression levels of VEGF of the cryoablation plus Endostar group were significantly lower than those of the other three groups (P<0.01). Statistical analysis revealed that a positive correlation existed between the express of VEGF and MVD. Conclusion: Endostar can obviously enhance the therapeutic efficacy of cryoablation on lung adenocarcinoma A549 in BALB/c nude mice. The underlying mechanisms may be the Endostar-inhibited angiogenesis through down-regulating the expression of VEGF, and the cooperative effect of Endostar and cryoablation on the promotion of tumor cell apoptosis. (authors)

  6. The mRNA and protein expression of folylpolyglutamate synthetase in methotrexate enantiomer-resistant A549 cell lines%信息动态

    2011-01-01

    Objective To study the expression of folylpolyglutamate synthetase ( FPGS ) in methotrexate ( MTX ) enantiomer-resistant A549 cell lines [ L-( + )-MTX and D-( - )-MTX ]. Methods The expression of FPGS on genetic and protein level was determined by FQ-PCR and Western blot in lung cancer A549 cells, and MTX enantiomer-resistant A549 cells [ L-( + )-MTX and D-( - )-MTX ], with the concentration of drug resistance was 15 μmol/L. Results The genetic expression level of FPGS was ( 0.80 ± 0. 09 ) and ( 2. 04 ± 0. 34 ) folds in L-( + )- MTX/A549 cells and D-( - )-MTX/A549 cells compared with lung cancer A549 cells, there was statistical difference between two groups ( P < 0.05 ). The protein expression level of FPGS was ( 0. 85 ± 0. 12 ) and( 1.62 ± 0. 24 ) folds in L-( + )-MTX/A549 cells and D-( - )-MTX/A549 cells compared with lung cancer A549 cells,there was statistical difference ( P < 0. 05 ). Conclusion The expression level of FPGS on genetic and protein level in drug resistant cells have been changed, and significant difference in two enantiomer-resistant cells are appeared.

  7. Inhibition of Oridonin on Human Lung Adenocarcinoma A549 Cells and Its Mechanisms%冬凌草甲素诱导人肺腺癌细胞株A549凋亡及其机制研究

    彭蕾; 顾振纶; 薛仁宇; 周颖; 蒋小岗; 郭次仪

    2010-01-01

    目的:探讨冬凌草甲素(oridnin)对人肺腺癌细胞株A549细胞凋亡的影响.方法:利用MTT法检测oridnin对A549细胞增殖作用的影响;Hoechst 33258染色观察给药后细胞形态改变;透射电镜观察给药后细胞超微结构改变;FITC-AnnexinV/PI双标记检测细胞凋亡率.结果:Hoechst 33258染色和透射电镜观察,oridonin给药后A549细胞出现空泡变性,染色质高度凝集;FTTC-AnnexinV/PI双标记检测oridnin(25,50,100μmol/L)作用细胞48 h后凋亡率分别为1.5%,6.2%,59.7%.结论:Oridonin对A549细胞具有抑制增殖和诱导凋亡作用.

  8. The Effects of Nimesulide Combined with Cisplatin on Lung Cancer

    邢丽华; 张珍祥; 徐永健; 张惠兰; 刘剑波

    2004-01-01

    To study the effects of cyclooxygenase 2 selective inhibitor Nimesulide (NIM) combined with Cisplatin (DDP) on human lung cancer and the possible mechanisms, the proliferation and apoptosis of human lung cancer cell line A549 were evaluated by MTT reduction assay and flow cytometry respectively. The inhibitory effect on neoplasia in vivo was tested on nude mice subcutaneously implanted tumor. Our results showed that NIM and DDP could inhibit A549 cell proliferation in a concentration dependent pattern; this action was enhanced when NIM (25 μmol/L) was given in combination with DDP and they worked in a synergistic or additive pattern as DDP concentration ≥ 1 μg/ml. NIM and DDP could induce A549 cells apoptosis and the action was augmented when used in combination (P<0.01). NIM and DDP could inhibit the growth of subcutaneously implan ted tumors on nude mice (P<0.05,P<0.01) and the inhibitory rate of NIM combined with DDP was significantly higher than that of NIM orDDPgroup (P<0.01, P<0.01). It is concluded that combined usc of NIM and DDP has significant synergistic antitumor effects on lung cancer cell line A549 and in animals in vivo. The synergy may be achieved by growth inhibition and apoptosis induction.

  9. Separation of an aqueous extract Inonotus obliquus (Chaga). A novel look at the efficiency of its influence on proliferation of A549 human lung carcinoma cells.

    Mazurkiewicz, Witold; Rydel, Katarzyna; Pogocki, Dariusz; Lemieszek, Marta Kinga; Langner, Ewa; Rzeski, Wojciech

    2010-01-01

    Aqueous extract of Inonotus obliquus was hydrolyzed in dilute hydrochloric acid. The products were extracted applying organic solvents, and separated chromatographically on a silica gel-packed column. Eluted fractions were analyzed by means of GC-MS. The presence of hydrocarbons, alcohols, phenols and various carbonyl compounds in analyzed fractions has been detected and quantified. Preliminarily experiments on the influence of certain separated samples on the proliferation of A549 human lung carcinoma cells were performed. Therefore, we hypothesize that the major antiproliferative effects are related to the presence of benzaldehyde, which is a benzyl alcohol metabolite formed in situ in the cells culture with the yield moderated by the presence of trace amounts of "high molecular mass compounds". PMID:20635536

  10. miR-155在肺腺癌 A549细胞侵袭和转移中的作用%Role of miR-155 in invasion and metastasis of lung adenocarcinoma A549 cells

    程田力; 胡成平; 李敏; 顾其华; 安健

    2016-01-01

    Objective To investigate the role and mechanism of miR-155 in invasion and metastasis of lung adenocarcinoma A549 cells.Methods Real-time PCR and fluorescence in situ hybridization were used to detect the miR-155 expression in patients′lung adenocarcinoma and adjacent tissue and lymph nodes.Scratch test and Transwell migration assay were used to assess the effect of miR-155 on the A549 cell migration and invasion capability.Bioinformatics software was used to predict the target genes of miR-155, and using luciferase to assay the target gene.Western blot and real-time PCR were performed to confirm the role of miR-155 expression in the regulation of target gene PTEN.Results The real-time quantitative PCR showed that the miR-155 expression levels in adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were 4.1±0.5, 9.6±3.1 and 7.8±2.2, respectively.The in situ hybridization showed that the expression rates of miR-155 in the adjacent normal tissue, lung adenocarcinoma and metastatic lymph nodes were (23.2±15.3)%, (75.4±20.2 )% and (60.4±25.1)%,respectively.The Scratch assay showed that the wound healing rates in the miR-155 mimics group, miR-155 mimics NC group, miR-155 inhibitor group and miR-155 inhibitor NC group at 24 h were (43.2±2.2)%, (21.3±4.2)%, (24.3±5.3)%, and (35.2± 5.1)%, and that at 48 h were (75.2±4.5)%, (52.6±5.2)%, (39.4±4.2)%, and( 51.5±4.3)%, respectively.Dual luciferase reporter gene assay showed that the value of the luciferase in the miR -155 mimics group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 4.7±0.5 and 7.3±0.7, and the miR-155 mimics luciferase values of the control group co-transfected with PTEN 3′UTR-containing wild-type and mutant plasmids were 7.8±0.9 and 7.5±0.8, respectively.The real-time quantitative fluorescence PCR showed that the relative expression of PTEN protein in the miR-155 mimics group, miR-155 mimics control group, miR-155 mimics inhibitor group