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Bmi1 基因沉默抑制人肝癌细胞的增殖和侵袭能力,并增加其对 5-氟尿嘧啶的敏感性。

Bmi1 gene silencing inhibits the proliferation and invasiveness of human hepatocellular carcinoma cells and increases their sensitivity to 5-fluorouracil.

机构信息

Department of Hepato-Pancreato-Biliary Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, Guangdong Province, PR China.

出版信息

Oncol Rep. 2013 Mar;29(3):967-74. doi: 10.3892/or.2012.2189. Epub 2012 Dec 14.

Abstract

The Bmi1 gene has been reported to play important roles in cancer initiation and progression. The aim of this study was to investigate the effects of RNA interference (RNAi)-mediated silencing of Bmi1 gene expression on the proliferation and invasiveness of hepatocellular carcinoma (HCC) cells and on the efficacy of chemotherapy in HCC patients. The Bmi1 gene was silenced by Bmi1-siRNA (small interfering RNA) in the human HCC cell lines HepG2 and Bel-7402, and the gene expression levels were assayed by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting. The proliferation and migration of Bmi1-silenced tumor cells and their sensitivity to 5-FU treatment were determined by Cell Counting Kit-8 (CCK-8), transwell assays and 4',6-diamidino-2-phenylindole (DAPI) staining and flow cytometry, respectively. Bmi1-siRNA inhibited the Bmi1 expression at both the mRNA and protein levels in HCC cells. Proliferation and migration of HCC cells treated with Bmi1-siRNA was significantly lower compared to that of the control cells. Moreover, Bmi1 gene silencing increased the percentage of apoptotic cells treated by 5-FU and decreased the IC50 values of 5-FU to a greater extent. Downregulation of the Bmi1 gene by RNAi can inhibit the proliferation and invasivesness of HCC cells and increase their sensitivity to 5-FU treatment.

摘要

Bmi1 基因已被报道在癌症的发生和进展中发挥重要作用。本研究旨在探讨 RNA 干扰(RNAi)介导的 Bmi1 基因表达沉默对肝癌(HCC)细胞增殖和侵袭能力以及 HCC 患者化疗效果的影响。通过 Bmi1-siRNA(小干扰 RNA)沉默人肝癌细胞系 HepG2 和 Bel-7402 中的 Bmi1 基因,并通过实时定量逆转录聚合酶链反应(qRT-PCR)和 Western blot 检测基因表达水平。通过细胞计数试剂盒-8(CCK-8)、Transwell 测定和 4',6-二脒基-2-苯基吲哚(DAPI)染色和流式细胞术分别测定 Bmi1 沉默肿瘤细胞的增殖和迁移及其对 5-FU 治疗的敏感性。Bmi1-siRNA 抑制 HCC 细胞中 Bmi1 的 mRNA 和蛋白水平表达。与对照组细胞相比,用 Bmi1-siRNA 处理的 HCC 细胞的增殖和迁移明显降低。此外,Bmi1 基因沉默使经 5-FU 处理的凋亡细胞比例增加,并且更显著地降低 5-FU 的 IC50 值。RNAi 下调 Bmi1 基因可抑制 HCC 细胞的增殖和侵袭能力,并增加其对 5-FU 治疗的敏感性。

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