Enhanced autophagic flux by endoplasmic reticulum stress in human hepatocellular carcinoma cells contributes to the maintenance of cell viability.-猫眼课题宝

DOI：

10.3892/or.2013.2474

发表时间：

2013-07

期刊：

Oncology reports

影响因子：

4.2

通讯作者：

中科院分区：

医学3区

文献类型：

作者：

Tai Ma;Yuan-yuan Li;Jie Zhu;Lu-lu Fan;Wei-Dong Du;Chang-hao Wu;G. Sun;Jia-bin Li

内质网应激和自噬是真核生物重要的适应性反应。本研究旨在探讨内质网应激下肝细胞癌细胞的自噬反应以及自噬对细胞存活和死亡的影响。用衣霉素刺激人肝癌细胞系HepG2诱导内质网应激。用细胞计数试剂盒检测细胞存活率。用透射电子显微镜观察到自噬小室的堆积。Western blotting检测ER和自噬相关蛋白的表达。用微管相关蛋白1-轻链3(MAP1-LC3)翻转实验评价氯喹抑制溶酶体时的自噬通量。在内质网应激下，HepG2细胞表现出显著的自噬小体积累，并增加了LC_3-I向LC_3-II的转化，并通过LC_3周转实验检测到自噬通量增加。3-甲基腺嘌呤抑制自噬促进了内质网应激相关的细胞死亡。我们的结论是，内质网应激增强了HepG2细胞的自噬通量，这可能有助于维持细胞的活力。

Endoplasmic reticulum (ER) stress and autophagy are important adaptive responses in eukaryotes. The aim of this study was to investigate the autophagic responses in hepatocellular carcinoma (HCC) cells under ER stress and the effect of autophagy on cell survival and death. The human HCC cell line HepG2 was stimulated with tunicamycin to induce ER stress. Cell viability was detected using the Cell Counting Kit-8. The accumulation of autophagic compartments was observed using transmission electron microscopy. The expression of ER and autophagy-related proteins was assessed by western blotting. Autophagic flux was assessed by microtubule-associated protein 1-light chain 3 (MAP1-LC3) turnover assay in the presence of chloroquine to inhibit lysosomes. HepG2 cells subjected to the ER stress presented a significant accumulation of autophagosomes and increased conversion of LC3-I to LC3-II as well as enhanced autophagic flux as detected by the LC3 turnover assay. Inhibition of autophagy with 3-methyladenine facilitated ER stress-related cell death. We conclude that ER stress enhances the autophagic flux in HepG2 cells, which may contribute to the maintenance of cell viability.

Enhanced autophagic flux by endoplasmic reticulum stress in human hepatocellular carcinoma cells contributes to the maintenance of cell viability.

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