Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line
Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line
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DOI：
10.7314/apjcp.2013.14.10.5849
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发表时间：
2013-01-01
影响因子：
--
通讯作者：
Chen, Wei
中科院分区：
文献类型：
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作者：
Du, Peng;Cao, Hua;Chen, Wei
背景：Bcl-2表达被抑制后，增殖性癌细胞可诱导凋亡。本研究集中于ABT 737激活的自噬对表阿霉素抗肿瘤作用的影响。方法：MTT法检测ABT 737对HepG 2肝癌细胞株的细胞毒作用，流式细胞仪检测细胞凋亡。荧光显微镜下测定线粒体膜电位。单丹酰尸胺（MDC）染色用于检测自噬的激活。Western blotting检测凋亡或自噬相关蛋白p53、p62、LC 3和Beclin 1的表达。结果：ABT 737和表阿霉素对HepG 2细胞的生长抑制作用呈剂量和时间依赖性。ABT 737和表柔比星单独使用均可诱导细胞凋亡，线粒体膜电位降低，凋亡蛋白表达增加。当用ABT 373和表阿霉素共同处理HepG 2细胞时，检测到凋亡进一步增加。此外，我们的研究结果表明，ABT 373或表阿霉素可以激活细胞自噬，增加自噬体的形成，增加自噬相关蛋白的表达和LC 3荧光斑点。结论：ABT 737通过凋亡和自噬机制影响癌细胞，并且ABT 737可以通过激活自噬和诱导凋亡来增强表阿霉素对HepG 2细胞的作用。
Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose-and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.