Wu Rong-Shou, Wu Lin-Quan, Li Ke-Hao, Li En-Liang, Feng Qian, Zhang Jing-Ling
Department of Hepatobiliary Surgery, Second Affiliated Hospital of Nanchang University; First Affiliated Hospital of Nanchang University, Nanchang 330006, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2016 Feb;36(2):244-9.
To investigate the abnormal expression of interferon-induced transmembrane protein 3 (IFITM3) in hepatocellular carcinoma (HCC) and the effect of IFITM3 knock-down on the biological behaviors of hepatocellular carcinoma HepG2 cells.
Western blot analysis and immunohistochemical staining were used to determine the expression of IFITM3 protein in 60 HCC samples and paired adjacent tissues. A small interfering RNA fragments of IFITM3 (IFITM3 siRNA) was transiently transfected into HepG2 cells and expressions of IFITM3 at mRNA and protein levels were examined by qRT-PCR and Western blotting. The changes in the proliferation of the transfected cells were determined using cell counting kit 8 (CCK8) assay, and the cell invasion and migration were tested using Transwell assay and wound-healing assay.
Compared with the adjacent tissues, HCC tissues expressed significantly higher levels of IFITM3. In HepG2 cells, transfection with IFITM3 siRNA resulted in significant down-regulation of IFITM3 expression at both the protein and mRNA levels and obviously suppressed cell proliferation, invasion, and migration ability as compared with the cells transfected with scrambled siRNA and control cells (P<0.05).
IFITM3, which is overexpressed in HCC, plays a vital role in the proliferation and invasion of HCC cells and may serve as a potential target for gene therapy of HCC.
探讨干扰素诱导跨膜蛋白3(IFITM3)在肝细胞癌(HCC)中的异常表达以及IFITM3基因敲低对肝癌HepG2细胞生物学行为的影响。
采用蛋白质免疫印迹分析和免疫组织化学染色法检测60例HCC组织及其配对癌旁组织中IFITM3蛋白的表达。将IFITM3的小干扰RNA片段(IFITM3 siRNA)瞬时转染至HepG2细胞,采用实时荧光定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测IFITM3在mRNA和蛋白水平的表达。采用细胞计数试剂盒8(CCK8)法检测转染后细胞增殖的变化,采用Transwell小室实验和划痕实验检测细胞侵袭和迁移能力。
与癌旁组织相比,HCC组织中IFITM3表达显著上调。在HepG2细胞中,转染IFITM3 siRNA后,IFITM3在蛋白和mRNA水平均显著下调,与转染乱序siRNA的细胞和对照细胞相比,细胞增殖、侵袭及迁移能力明显受到抑制(P<0.05)。
IFITM3在HCC中高表达,对HCC细胞的增殖和侵袭起着重要作用,可能成为HCC基因治疗的潜在靶点。