南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (02): 244-.

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沉默干扰素诱导跨膜蛋白3可抑制肝癌细胞HepG2增殖和侵袭

吴荣寿,邬林泉,李科浩,李恩亮,冯潜,张惊玲   

  • 出版日期:2016-02-20 发布日期:2016-02-20

Interferon-induced transmembrane protein 3 knock-down inhibits proliferation and
invasion of hepatocellular carcinoma HepG2 cells in vitro

  • Online:2016-02-20 Published:2016-02-20

摘要: 目的研究干扰素诱导跨膜蛋白3(IFITM3)在原发性肝癌中的异常表达,探讨沉默IFITM3表达对肝癌细胞系HepG2生
物学特征的影响。方法通过Western blot和免疫组织化学染色法检测60例肝癌组织及对应癌旁组织中IFITM3的表达情况及
相关性;构建IFITM3小干扰RNA片段(IFITM3 si-RNA),采用脂质体介导转染肝癌细胞(HepG2细胞系),同时设置无义序列组
和空白对照组,实时荧光定量PCR检测转染后IFITM3 mRNA的表达;CCK8(cell counting kit 8)检测转染后细胞增殖能力;
Western blot检测IFITM3在蛋白水平的表达;Transwell实验和划痕实验检测细胞侵袭和迁移能力的变化。结果和癌旁组织相
比,IFITM3在肝癌组织中的明显高表达。与无义序列组和空白对照组比较,IFITM3 si-RNA转染组细胞IFITM3表达和细胞增
值率降低,穿膜细胞数也明显减少,差异均有统计学意义(P<0.05)。划痕实验也表明迁移能力降低。结论IFITM3在原发性肝
癌中高表达;IFITM3在肝癌细胞增殖和侵袭过程中发挥重要作用,有望成为原发性肝癌基因治疗的侯选靶点。

Abstract: Objective 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. Methods 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. Results
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). Conclusion 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.