Wnt信号通路在HepG2细胞株及其克隆形成细胞中表达的异质性  

Heterogeneous expression of Wnt signal molecules in parental HepG2 cells and HepG2 colony-forming cells

作　　者：王宝松[1] 魏德全[1] 于树娜[1] 姜栋栋[1] 赵金岭[1] 史才兴[1] 蒋吉英[1] 

机构地区：[1]潍坊医学院解剖学教研室,山东省潍坊市261053

出　　处：《世界华人消化杂志》2011年第3期284-288,共5页World Chinese Journal of Digestology

基　　金：山东省"泰山学者"建设工程专项基金资助项目;山东省科技攻关计划项目基金资助项目;No.2006GG3202016~~

摘　　要：目的:研究Wnt信号传导通路的关键因子β-catenin和COX-2在肝癌细胞株HepG2及其克隆形成细胞中的表达,探讨Wnt信号传导通路在不同增殖能力细胞中表达的异质性.方法:以HepG2细胞为研究对象,采用软琼脂克隆形成实验筛选克隆形成细胞,应用RT-PCR、免疫化学和Westernblot等技术,检测Wnt信号传导通路的关键因子β-catenin和COX-2在肝癌细胞株HepG2及其克隆形成细胞中的表达.结果:β-cateninmRNA和蛋白质在克隆形成细胞中的表达水平高于HepG2细胞(0.905vs0.549;1.021vs0.700;均P<0.05).β-catenin的阳性信号在HepG2细胞中主要分布于细胞质,克隆形成细胞中主要分布于细胞核.COX-2mRNA和蛋白质在HepG2细胞中的表达高于克隆形成细胞(0.857vs0.527;0.731vs0.434;均P<0.05).大多数HepG2细胞中表达COX-2蛋白,阳性反应沉淀物主要位于细胞质,少数克隆形成细胞中有COX-2蛋白弱表达.结论:Wnt信号传导途径的关键因子β-catenin和COX-2与肝癌分化程度相关.β-catenin和COX-2等Wnt信号传导通路的关键因子在肝癌细胞HepG2及其克隆形成细胞的分布模式支持克隆形成细胞具有肝癌干细胞的特征.Abstract AIM： To investigate the heterogeneous expression of Wnt signal molecules in HepG2 cells with different proliferative ability. METHODS： The soft agar colony formation assay was used to separate HepG2 colony-forming cells. The mRNA expression and protein expression and distribution of β-catenin and COX-2 in parental HepG2 cells and HepG2 colony- forming cells were detected by RT-PCR, Western blotting and immunochemistry, respectively. RESULTS： The expression levels of β-catenin mRNA and protein in HepG2 colony-forming cells were significantly higher than those in parental HepG2 cells （0.905 vs 0.549; 1.021 vs 0.700; both P 〈 0.05）. β-catenin was mainly localized in the cytoplasm in parental HepG2 cells and in the nucleus in HepG2 colony-forming cells. The expression levels of COX-2 mRNA and protein in parental HepG2 cells were significantly higher than those in HepG2 colony-forming cells （0.857 vs 0.527; 0.731 vs 0.434; both P 〈 0.05）. Immunochemistry analysis showed that most HepG2 cells were positive for COX-2. CONCLUSION： β-catenin and COX-2 may be closely related to the degree of differentiation of hepatocellular carcinoma. The expression patterns of β-catenin and COX-2 in parental HepG2 cells and HepG2 colony-forming cells imply that HepG2 colony-forming cells have the characteristics of liver cancer stem cells.

关 键 词：WNT Β-CATENIN COX-2 软琼脂克隆 肝癌 HEPG2 

分 类 号：R735.7[医药卫生—肿瘤]