GP73基因沉默对肝癌细胞增殖和凋亡的影响
Effect of GP73 silencing on proliferation and apoptosis in hepatocellular cancer.
作者信息
Zhang Yu-Long, Zhang You-Cheng, Han Wei, Li Yu-Min, Wang Geng-Nian, Yuan Shao, Wei Feng-Xian, Wang Jia-Feng, Jiang Jian-Jun, Zhang Ya-Wu
机构信息
Yu-Long Zhang, You-Cheng Zhang, Wei Han, Yu-Min Li, Geng-Nian Wang, Shao Yuan, Feng-Xian Wei, Jia-Feng Wang, Jian-Jun Jiang, Ya-Wu Zhang, Department of General Surgery, Lanzhou University Second Hospital, Lanzhou 730030, Gansu Province, China.
出版信息
World J Gastroenterol. 2014 Aug 28;20(32):11287-96. doi: 10.3748/wjg.v20.i32.11287.
AIM
To investigate the roles of Golgi protein (GP) 73 in the regulation of cell proliferation and apoptosis.
METHODS
Stealth RNAi targeting GP73 gene sequence was used to silence its expression in Hep G2 cells and Bel7402 cells. Stealth RNAi effects were assessed by reverse transcriptase polymerase chain reaction and ELISA. Cell proliferation assay and cell cycle analysis were assessed by MTT assay and flow cytometry. Apoptosis was assessed by flow cytometry and transmission electron microscopy. Apoptosis-related proteins were assessed by western immunoblot analysis.
RESULTS
Stealth RNAi targeting GP73 gene sequence markedly reduced the expression of GP73 gene. The reduction of GP73 in Hep G2 cells and Bel7402 cells inhibited cell proliferation and induced apoptosis, however, terminal apoptosis occurred in Hep G2 cells, but early apoptosis occurred in Bel7402 cells. Reduced expression of GP73 gene might lead to a reduction in Bcl-2/Bax ratio, an increase in cytochrome c, but a reduction in capase-3.
CONCLUSION
GP73 might play an important role in proliferation and apoptosis in hepatocellular carcinoma cells.
目的
探讨高尔基体蛋白（GP）73在细胞增殖和凋亡调控中的作用。
方法
采用针对GP73基因序列的Stealth RNAi沉默其在肝癌细胞Hep G2和Bel7402中的表达。通过逆转录聚合酶链反应和酶联免疫吸附测定评估Stealth RNAi的效果。采用MTT法和流式细胞术评估细胞增殖及细胞周期分析。通过流式细胞术和透射电子显微镜评估细胞凋亡。通过蛋白质免疫印迹分析评估凋亡相关蛋白。
结果
针对GP73基因序列的Stealth RNAi显著降低了GP73基因的表达。Hep G2细胞和Bel7402细胞中GP73表达降低抑制了细胞增殖并诱导了凋亡，然而，Hep G2细胞发生终末凋亡，而Bel7402细胞发生早期凋亡。GP73基因表达降低可能导致Bcl-2/Bax比值降低、细胞色素c增加，但caspase-3降低。
结论
GP73可能在肝癌细胞的增殖和凋亡中发挥重要作用。
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