Influence of MMP-9 antiseme oligonudeotide on proliferation and apoptosis of human lung adenocarcinom A549 cells / 中国医师杂志

Objective To approach the effects of MMP-9 antisensc oligonucleotide(ASODN)on human lung adcnocarcinoma(A549) cell apoptosis and proliferation capability.Methods MTT.was used to analyze the effect that MMP-9 transfection on A549 cell growth.Flow cytometry was used to analyze the ratio of cell proliferation and apoptosis.RT-PCR WaS used to detect the expression of MMP-9mRNA and Western blot was used to detect the changes of MMP-9 protein.Results In some degree,MMP-9 ASODN that inhibited the survival rate of the A549 cell presented concentration and time dependence manner.The best dependence concentration of ASODN was 600nmol/L for 24 hour.After trasnfection of MMP-9ASODN to A549 cell.the perceive of A549 apoptosis cell was significandy higher than that in control group(P＜0.01).When the antisense oligonucleotides concentration is 600nmol/L and the action time is 48 hours,the relative expression level of MMP-9mRNA and MMP-9 protein are obviously less than that in control group(P＜0.01).Conclusion MMP-9ASODN may down regulate the expression of MMP-9mRNA and MMP-9 protein,effectively inhibit the proliferation of A549 cell and promote the apoptosis of A549 cell.

Relationship between ERCC1 expression and cisplatin intervention in human lung adenocarcinoma cell lines / 中国肺癌杂志

<p><b>BACKGROUND</b>The excision repair cross-complementing gene 1 (ERCC1), which is important in the repair of cisplatin-DNA adducts, is reported to be related to cisplatin resistance in tumor cells. The aim of this study is to investigate the influence of low-dose cisplatin on expression of ERCC1 gene and to confirm the correlation between ERCC1 and cisplatin resistance in human lung adenocarcinoma cell lines.</p><p><b>METHODS</b>A549 and A549/DDP cell lines were treated with 10 μmol/L cisplatin for 12, 24, 48 and 72 h, or treated with 5, 10, 20, 40 μmol/L cisplatin for 24 h respectively. Then the expression of ERCC1 mRNA and protein was measured by RT-PCR and immunocytohistology SABC assay respectively. The resistance of A549/DDP cells was measured by MTT assay.</p><p><b>RESULTS</b>After treating with 10 μmol/L cisplatin for 12 h, up-regulation of ERCC1 mRNA and protein was observed in A549 cells, then reached the peak levels in 72 h group. After treating with 5 μmol/L cisplatin for 24 h, up-regulation of ERCC1 mRNA and protein was observed in A549 cells, and when treated with 20 μmol/L cisplatin for 24 h, the ERCC1 mRNA and protein reached the peak levels. Comparing with the parental cells, ERCC1 expression increased obviously in A549/DDP cells, which were established by continuous low-dose cisplatin treatment.</p><p><b>CONCLUSIONS</b>Up-regulation of ERCC1 expression can be induced by low-dose cisplatin in human lung adenocarcinoma cell line A549, and ECRR1 may play roles in cisplatin resistance.</p>

Study on apoptosis of human lung adenocarcinoma cell line A549/DDP induced by ginsenoside Rh₂ in vitro / 中国肺癌杂志

<p><b>BACKGROUND</b>Lung cancer is one of the leading causes of cancer-related death in mankind. To exploit antitumor drug from plant has been a highlight at home and abroad. The aim of this study is to investigate the apoptosis of human lung adenocarcinoma cell line A549/DDP induced by ginsenoside Rh₂ (G-Rh₂) and to explore its possible molecular mechanism.</p><p><b>METHODS</b>The growth inhibition effect of G-Rh₂ on A549/DDP cells was evaluated by MTT assay. Cell cycle analysis, apoptosis index and tumor related gene expression were detected by flow cytometry. The changes of sApo-1/Fas level in the cell culture supernatant were determined by ELISA method.</p><p><b>RESULTS</b>(1) G-Rh₂ significantly inhibited the growth of A549/DDP cells in a dose-time-de-pendent manner. (2) After 24 hours' treatment with G-Rh₂, apoptosis index of trial group was significantly higher than that of control group (P < 0.001). The proportion of cells in G0/G1 phase in trial group was much higher than that in control group (P < 0.01), while proportion in S phase in trial group was markedly lower than that in control group (P < 0.01). There was no significant difference in proportion in G2/M phase between trial group and control group (P > 0.05). (3) The positive expression rate of p53 and Fas in trial group was significantly higher than that in control group (P < 0.01, P < 0.001), while the positive expression rate of Bcl-2 in trial group was significantly lower than that in control group (P < 0.001). (4) The level of sApo-1/Fas in A549/DDP cell culture supernatant in trial group was remarkably lower than that in control group (P < 0.05).</p><p><b>CONCLUSIONS</b>G-Rh₂ can induce the apoptosis of A549/DDP cells. Its molecular mechanism may be up-regulating expression of p53 and Fas and down-regulating expression of Bcl-2.</p>