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Effects of SIRT6 Gene Silencing on the Apoptosis in Human Hepatocellular Carcinoma Cells and Its Potential Mechanisms
Zhou Hongzhong, Tao Nana, Chen Xiang, Ren Jihua, Li Wanyu, Liu Bo, Chen Juan*
Key Laboratory of Molecular Biology on Infection Diseases of Ministry of Education,Chongqing Medical University, Chongqing 400016, China
Abstract: This study investigated the effects of lentivirus-mediated silent information regulator 6 (SIRT6) knock-down on cell apoptosis in hepatocellular carcinoma (HCC) cells and its potential mechanisms. The expression of SIRT6 gene in HCC cell lines (SK-Hep-1， Huh-7， PLC/PRF/5， HepG2) and immortalized human liver cell line (MIHA) was detected by reverse-transcription polymerase chain reaction (RT-PCR) and Western blot， respectively.SIRT6 gene was silenced by lentivirus-mediated shRNA interference technology and its efficiency of SIRT6 gene silencing was detected by RT-PCR and Western blot. Effects of SIRT6 gene silencing on apoptosis in SK-Hep-1 and Huh-7 cells were analyzed by flow cytometry. The mRNA and protein levels of IAPs (inhibitor of apoptosis proteins) family were analyzed by RT-PCR and Western blot， respectively. Effects of XIAP (X-linked inhibitor of apoptosis protein gene) on the induction of apoptosis in SIRT6-depleted SK-Hep-1 cells were analyzed by flow cytometry. Our results showed that SIRT6 was over-expressed in HCC cells. Lentivirus-mediated shRNA interference down-regulated the protein level of SIRT6 in the HCC cells. Knockdown of SIRT6 gene induced the apoptosis of HCC cells and down-regulated the mRNA and protein levels of XIAP. XIAP over-expression significantly abolished the induction of apoptosis in SIRT6-depleted cells. Together， these data indicated that SIRT6 gene silencing might induce cell apoptosis in human hepatocellular carcinoma cells by regulating XIAP protein level.
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