Caveolin-1真核表达载体的构建及其在Hepa1-6细胞中的表达  

Construction and expression of caveolin-1 expression vector in hepa1-6 cells

作　　者：汪淑晶[1] 汪晓敏[1] 师伟[1] 张嘉宁[1] 

机构地区：[1]大连医科大学生物化学与分子生物学教研室糖生物学研究所,大连116044

出　　处：《生物学杂志》2011年第6期26-30,共5页Journal of Biology

基　　金：国家自然科学基金资助项目(31000372,30970648)

摘　　要：Caveolin-1在不同肿瘤中发挥作用不同,既发挥抑癌基因样作用又发挥癌基因样作用。旨在分析caveolin-1在小鼠肝癌细胞系中的表达情况及建立稳定表达外源caveolin-1的Hepa1-6细胞。利用RT-PCR和Western-blot方法检测caveolin-1在小鼠肝癌H22、Hca-F和Hepa1-6细胞中的表达;通过分子克隆构建小鼠caveolin-1 cDNA真核表达载体,利用脂质体转染等方法建立稳定表达外源caveolin-1的Hepa1-6细胞株;通过RT-PCR、Western-blot、免疫细胞化学等方法鉴定其稳定表达细胞株。结果显示,caveolin-1在Hepa1-6细胞中表达呈阴性,在H22和Hca-F中高表达;成功获得小鼠caveolin-1 cDNA真核表达载体pEGFP-N2/Cav-1,筛选并鉴定出高表达外源caveolin-1的Hepa1-6稳定细胞株C1和C4,为进一步分析caveolin-1在肝癌中所发挥的作用奠定了一定的研究基础。Caveolin-1 plays different roles in different kinds of tumors. Caveoiln-1 may act as tumor-suppressor, as well as tumor-promoter. This study aims to analyze the expression of caveolin-1 in mouse hepatoma cell lines and construct stable hepal-6 cells expressing exogenous eaveolin-1. The expression of caveolin-1 in mouse hepatoma cell lines （H22,Hca-F and Hepal-6）was investigated by RT-PCR and Westeru-blot assays; mouse caveolin-I eDNA expression vector was constructed by molecular clone, and. stable hepal-6 cells transfected with caveolin-1 gene were obtained through transfection and screening. Results showed that the expression levels of caveolin-1 were undetectable in hepal-6 cells, and upregulated in H22 and Hca-F cells. Mouse caveolin-1 eDNA expression vector and stable transfected hepal-6 cells were generated successfully. The stable hepal-6 cells transfected with caveolin-1 will lay a foundation for ~rther studies of caveolin-1 function.

关 键 词：CAVEOLIN-1 分子克隆 转染 肝癌 

分 类 号：Q78[生物学—分子生物学]