论文部分内容阅读
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 tumors, both play a role of tumor suppressor gene-like and play an oncogene-like role. The aim of this study was to analyze the expression of caveolin-1 in mouse hepatocellular carcinoma cell lines and to establish Hepa 1-6 cells stably expressing exogenous caveolin-1. The expression of caveolin-1 in H22, Hca-F and Hepa1-6 cells was detected by RT-PCR and Western-blot. The mouse caveolin-1 cDNA eukaryotic expression vector was constructed by molecular cloning. Transfection and other methods to establish a stable expression of exogenous caveolin-1 Hepa1-6 cell lines; RT-PCR, Western-blot, immunocytochemistry and other methods to identify stable expression of cell lines. The results showed that caveolin-1 was negatively expressed in Hepa1-6 cells and highly expressed in H22 and Hca-F. Mouse caveolin-1 cDNA eukaryotic expression vector pEGFP-N2 / Cav-1 was successfully obtained and screened and identified as high The expression of caveolin-1 exogenous Hepa1-6 stable cell lines C1 and C4, for further analysis of the role of caveolin-1 in liver cancer has laid a foundation for the study.