AGR2调控IFITM3的表达促进肝癌细胞的增殖

AGR2 regulates IFITM3 expression to promote hepatocellular carcinoma cell proliferation

目的:探讨重组人前梯度同源蛋白2(anterior gradient homolog 2,AGR2)的表达对肝癌细胞增殖能力的影响及其可能的作用机制.方法:通过实时荧光定量PCR、Western blot、免疫组织化学检测40例肝癌患者癌组织和对应癌旁组织中AGR2基因、干扰素诱导跨膜蛋白3(interferon-induced transmembrane protein 3,IFITM3)的表达情况,构建AGR2的过表达质粒pcDNA3.1-AGR2,并将其转染至肝癌细胞中,运用实时荧光定量PCR和蛋白质印迹法分别检测AGR2 mRNA和蛋白的表达,CCK-8(cell counting kit 8)法检测细胞的增殖能力,利用流式细胞技术检测细胞凋亡,蛋白质印迹法检测IFITM3的表达.结果:在原发性肝癌组织中AGR2,IFITM3呈现高表达;成功构建AGR2的过表达质粒pcDNA3.1-AGR2,成功构建稳定高表达AGR2的肝癌HepG2细胞.pcDNA3.1-AGR2转染组HepG2细胞中AGR2 mRNA和蛋白的表达水平明显高于阴性对照组和空白对照组.与阴性对照组比较,pcDNA3.1-AGR2转染组HepG2细胞的体外增殖能力和克隆形成能力明显升高,IFITM3蛋白的表达水平也随之升高.结论:上调AGR2可以增加IFITM3的表达,促进肝癌细胞的增殖.

AIM：To investigate the effect of anterior gradient homolog 2（AGR2） expression on the proliferation of hepatocellular carcinoma cells,and to explore the possible mechanism.METHODS：The expression of AGR2 and interferon-induced transmembrane protein3（IFITM3） in hepatocellular carcinoma and adjacent tissues was detected by fluorescence quantitative PCR and Western blot.The expression of AGR2 mRNA and protein was examined by real-time fluorescence quantitative PCR and Western blot after transfection with pcDNA3.1-AGR2.The cellular growth ability was examined by CCK-8 assay,and the colony formation ability was detected by colony formation assay.Flow cytometry assay was used to determine the apoptosis index.The expression level of IFITM3 protein was examined by Western blot.RESULTS：AGR2 and IFITM3 mRNA and protein expression in hepatocellular carcinoma tissues was significantly higher than that in the corresponding adjacent tissues.The pcDNA3.1-AGR2 was successfully constructed,and HepG2 cells with stable expression of AGR2 were established.The expression levels of AGR2 mRNA and protein in HepG2 cells after transfection with pcDNA3.1-AGR2 were higher than those in the negative control cells,and the cellular growth ability of HepG2 cells after transfection with pcDNA3.1-AGR2 was significantly increased.IFITM3 protein expression was also increased after transfection with pcDNA3.1-AGR2.CONCLUSION：Raised AGR2 expression can increase the expression of IFITM3 and promote the proliferation of hepatocellular cells.