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目的:探讨去甲基化制剂5-氮-2′-脱氧胞苷(5-Aza-CdR or DAC)和组蛋白去乙酰化酶抑制剂曲古抑菌素A(tri-chostatin A,TSA)对人子宫内膜腺癌Ishikawa和AN3细胞增殖和凋亡的影响。方法:台盼蓝拒染法观察药物对细胞生长曲线的影响;AnnexinⅤ染色法检测细胞凋亡;流式细胞仪分析细胞周期;Western blot检测凋亡信号通路中Caspase-8、Caspase-9活化及多聚ADP核糖聚合酶(PARP)裂解情况。结果:DAC和TSA对Ishikawa和AN3细胞均有时间依赖性的抑制作用,联用后抑制作用更强。单用DAC或TSA均可诱导细胞凋亡。单用DAC或TSA对细胞G1期无影响,单用TSA使G2/M期细胞比例增高,联合用药发生明显的G1期阻滞。Western blot检测表明,DAC和TSA诱导了Caspase-8、Caspase-9裂解活化及PARP裂解。结论:DAC与TSA协同可抑制细胞生长,并使细胞阻滞在G1期,且细胞凋亡发生的时相不同于单独用药组。
Objective: To investigate the effects of 5-Aza-CdR DAC and histone deacetylase inhibitor trichostatin A (TSA) Effects of Proliferation and Apoptosis on Human Endometrial Adenocarcinoma Ishikawa and AN3 Cells. Methods: The effect of drugs on cell growth curve was observed by trypan blue exclusion staining. Cell apoptosis was detected by Annexin Ⅴ staining. Cell cycle was analyzed by flow cytometry. The activation of Caspase-8 and Caspase-9 in apoptosis signal pathway was detected by Western blotting. Poly ADP ribose polymerase (PARP) cleavage. Results: Both DAC and TSA inhibited the proliferation of Ishikawa and AN3 cells in a time-dependent manner. Both DAC and TSA alone induce apoptosis. Single cell DAC or TSA had no effect on cell G1 phase. With TSA alone, the proportion of cells in G2 / M phase was increased, and G1 phase arrest was observed in combination therapy. Western blot showed that DAC and TSA induced the cleavage of Caspase-8, Caspase-9 and PARP cleavage. Conclusion: The synergistic effect between DAC and TSA can inhibit the cell growth and block the cell cycle in G1 phase, and the phase of apoptosis is different from the single drug group.