Caveolin一1真核表达载体的构建及其在Hepal-6细胞中的表达

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-l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Hepal-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Hepal-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Hepal-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-l在Hepal-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Hepal-6稳定细胞株Cl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。

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

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.