肝癌组织与细胞中缺氧诱导因子-2α、HBx 表达的相关性研究

摘要

Objective Objective To investigate the relationship between hypoxia-inducible factor-2α( HIF-2α) and hepatitis B virus X protein (HBx) in hepatocellular carcinoma (HCC) tissues and cells.Methods The HCC Bel-7404 cell line was cultured and then was divided into 6 groups:groups A, B and C were transfected by 1.2, 0.6, and 0.3 μg pEGFP-HBx;the no-load transfection group was transfected by 1.2 μg pEGFP; the control group was only added lipido-some;and the non-transfection group was not added any lipidosome and plasmids .Real-time PCR and Western blotting were used to investigate the mRNA and protein expression of HIF-1αand HIF-2α.Immunohistochemistry was used to de-tect the expression of HIF-2αand HBx in 33 HCC tissues , and their possible relationships were analyzed by using SPSS 15.0 statistical software.Results The mRNA and protein expression of HIF-1αand HIF-2αin Bel-7404 cells:group A>group B >group C >no-load transfection group >control group >non-transfection group (all P<0.05).The ex-pression of HIF-1αand HIF-2αwas increased in Bel-7404 cells at both mRNA and protein levels in a dose-dependent man-ner after being transfected by HBx plasmid (all P<0.05).HIF-2αwas highly expressed in HCC tissues , but was not cor-related with the expression of HBx (P>0.05).Conclusion HBx may up-regulate the expression of HIF-2αand HIF-1αin HCC cells, but it is not correlated with HIF-2αexpression in HCC tissues .%目的：观察肝癌组织与肝癌细胞中缺氧诱导因子（ HIF）-2α与乙型肝炎病毒X蛋白（ HBx）表达的相关性。方法培养肝癌细胞系Bel-7404，将细胞分为6组：A、B、C组分别转染1．2、0．6、0．3μg pEGFP-HBx，空载体组转染1．2μg pEGFP，对照组仅加脂质体，未转染组不加脂质体及质粒。 Real-time PCR法检测各组细胞中的HIF-1α、HIF-2αmRNA，Western blot法检测其蛋白。采用免疫组化法检测33例肝癌组织中的HIF-2α、HBx，并进行相关性分析。结果 Bel-7404细胞中HIF-1α、HIF-2αmRNA、蛋白表达A组＞B组＞C组＞空载体组、对照组、未转染组（P均＜0．05），A、B、C组HIF-1α、HIF-2αmRNA及蛋白表达呈HBx剂量依赖性（P均＜0．05）。肝癌组织中HBx、HIF-2α表达增高，但二者表达无相关性（P＞0．05）。结论肝癌细胞中，HBx可上调HIF-2α、HIF-1α的表达，但在肝癌组织中HBx与HIF-2α表达无相关性。