HBx基因上调组织蛋白酶S对HepG2 细胞的影响  

Effects of up-regulation of cathepsin S by HBx gene on HepG2 cell

作　　者：张智[1] 孙兴[1] 肖艺[1] 方兆山 黄海[1] 吴曙粤[2] 徐静[3] 郭鹏[4] Zhang Zhi Guo Peng Sun Xing Xiao Yi Xu Jing Fang Zhaoshan Wu Shuyue Huang Hai(Department of Hepatobiliary and GlandSurgery, The First People＇s Hospital of Nanning, Nanning 530022, Chin)

机构地区：[1]南宁市第一人民医院肝胆腺体外科,广西南宁530022 [2]广西医科大学附属口腔医院口腔外科 [3]广西医科大学第一附属医院肝胆外科 [4]南宁市第一人民医院儿科,广西南宁530022

出　　处：《中华医学杂志》2017年第30期2357-2361,共5页National Medical Journal of China

基　　金：广西医科大学青年科学基金项目（GxMuYsF201542）;南宁市科技计划项目（20163130）;南宁市普通外科腹腔镜微创中心建设（20153012）

摘　　要：目的 检测乙型肝炎病毒e抗原（HBeAg）（-）、（＋）的肝细胞肝癌（HCC）组织乙肝病毒X基因（HBx）、组织蛋白酶S（Cat S）表达情况,进一步探讨HBx与Cat S相互作用对HepG2细胞的影响.方法 收集2015年1月至2016年5月间广西南宁市第一人民医院手术切除经病理证实的HCC标本70例,根据临床血清学检验结果将其分为HBeAg（-）和HBeAg（＋）两组.通过免疫组化检测Cat S、HBx在HCC组织和癌旁组织中表达情况,进一步构建pcDNA3.1-HBx质粒和对照质粒,瞬时转染HepG2细胞,收集细胞,用Western 印迹实验检测Cat S和HBx蛋白表达,通过MTT实验、划痕实验和Transwell实验观察细胞增殖情况.结果 HBeAg（＋）的HCC组织中Cat S的阳性率明显高于HBeAg（-）的HCC组织（52.67%±0.33%与41.23%±0.52%,P〈0.05）;在HBeAg（＋）的HCC组织HBx阳性表达率达92.89%,而在HBeAg（-）的HCC组织中未见阳性表达.与转染对照质粒的HepG2对照组相比,实验组细胞Cat S和HBx蛋白表达水平明显增加,细胞增殖、迁移（21.98%±1.69%与58.23%±1.47%）和侵袭（24.12%±1.15%与 64.25%±1.42%）明显增强,差异有统计学意义（P〈0.05）.结论 在HCC组织中HBx和Cat S的表达呈正相关,HBx基因可能通过上调Cat S基因的表达促进HepG2-HBx细胞增殖.Objective To explore the expression of cathepsin S （Cat S） and Hepatitis B virus X protein （HBx） in HBeAg（-） and HBeAg（＋） hepatocellular carcinoma （HCC）, and discuss the effects of Cat S and HBx interaction on HepG2 cell.Methods Seventy HCC tissue specimens were collected from the surgical resection which were confirmed by pathology in the Fifth Affiliated Hospital of Guangxi Medical University. The tissue samples were separated into two groups： HBeAg（-） group and HBeAg（＋） group according to the serology of HBeAg. The expression of Cat S and HBx in the para-carcinoma tissue and the HCC tissue was determined by immunohistochemical staining. The recombinant plasmid of pcDNA3.1-HBx and empty plasmid were constructed and transfected transiently into HepG2 cell. Cells were harvested, and Western blot assay was performed to detectthe protein expression of Cat S and HBx. The cell proliferation was measured by methyl thiazolyl tetrazolium （MTT） assay, wound healing assay and transwell migration assay.Results Immunohistochemical staining showed that Cat S expression was up-regulated in HBeAg（＋） HCC cancer tissues, compared with HBeAg（-）HCC cancer tissues （52.67%±0.33% vs 41.23%±0.52%, P〈0.05）. HBx expression was up-regulated in HBeAg（＋） HCC cancer tissues （92.89%）, but not in HBeAg（-）HCC cancer tissues. Compared with HepG2 control group, cells in HepG2-HBx group had significantly higher protein level of Cat S and HBx, more obvious proliferation and migration （21.98%±1.69% vs 58.23%±1.47%） and invasion （24.12%±1.15%vs 64.25%±1.42%） （all P〈0.05）.Conclusions The expression of HBx and Cat S had a linear positive correlation in liver tissues, and increased expression of HBx can promote the cell proliferation of HepG2-HBx cell line.

关 键 词：肝细胞癌 乙肝病毒X基因 组织蛋白酶S 

分 类 号：R735.7[医药卫生—肿瘤]