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目的探讨阿糖胞苷(Ara-C)对人肺腺癌细胞株A549的凋亡诱导作用及其机制。方法Ara-C体外作用于A549细胞,噻唑蓝还原法(MTT法)检测Ara-C对A549细胞增殖的抑制作用;Hoechst33258荧光染色观察细胞核形态学的变化;单细胞凝胶电泳技术(comet assay)测定A549细胞DNA的损伤程度;以Western blotting进一步证明A549细胞发生凋亡。结果Ara-C对A549细胞的增殖有明显抑制作用;观察到特异性的凋亡小体;Ara-C导致A549细胞发生DNA链断裂,并呈明显剂量依赖性增强:Western blotting显示caspase 8,9,3都不同程度被激活,多聚ADP核糖聚合酶(PARP)被剪切降解。结论揭示了Ara-C明显诱导A549细胞凋亡;细胞凋亡通路不仅通过线粒体途径,还通过膜受体途径,两条通路协同作用使凋亡信号增强;提示Ara-C对A549细胞有明显的化疗作用。
Objective To investigate the apoptosis-inducing effect of Ara-C on human lung adenocarcinoma cell line A549 and its mechanism. Methods A549 cells were treated with Ara-C in vitro. The inhibitory effect of Ara-C on the proliferation of A549 cells was detected by MTT assay. The morphological changes of nuclei were observed by Hoechst33258 staining. The single cell gel electrophoresis (comet assay) The degree of DNA damage in A549 cells was determined. Western blotting was used to confirm the apoptosis of A549 cells. Results Ara-C significantly inhibited the proliferation of A549 cells; specific apoptotic bodies were observed; Ara-C resulted in DNA strand breaks in A549 cells in a dose-dependent manner: Western blotting showed that caspase 8,9 , 3 are activated to varying degrees, poly ADP ribose polymerase (PARP) is shear-degraded. The results showed Ara-C significantly induced A549 cell apoptosis; apoptosis pathway not only through the mitochondrial pathway, but also through the membrane receptor pathway, the two pathways synergistic effect so that apoptosis signal enhanced; Ara-C on A549 cells were significantly Chemotherapy.