βig-h3在肝癌细胞系中差异表达的筛选和鉴定

Screening and identification of differential expression of βig-h3 in human hepatoma cell lines

目的:筛选并鉴定TGFβ诱导分子βig-h3基因在肝癌细胞系T7721和7721中的差异表达.方法:培养肝癌细胞系T7721(转染并稳定高表达HAb18G/CD147),7721(未转染HAb18G/CD147),提取总RNA,荧光交换标记(Cy5和Cy3)筛选信号转导相关基因芯片;RT-PCR检测基因芯片初步筛选出的差异分子(βig-h3)在肝癌细胞T7721和7721中的差异表达.结果:①经基因芯片差异分析,在高表达HAb18G/CD147的T7721细胞中TGFβ诱导分子βig-h3的mRNA表达明显上调,为7721细胞的4.4倍;而TGFβ1的mRNA表达无明显差别.②RT-PCR显示βig-h3mRNA在T7721中的表达水平明显高于7721(t=14.42,P<0.01).结论:经基因芯片筛选分析,发现HAb18G/CD147分子与TGFβ诱导分子βig-h3的表达成正相关,HAb18G/CD147的高表达诱导了βig-h3的表达增高,为进一步探讨在细胞内信号转导通路中HAb18G/CD147分子促进肝癌细胞侵袭和转移的作用机制奠定了基础.

AIM： To screen and identify the differential expression of TGFβ-induced molecule βig-h3 in hepatoma cell lines T7721 and 7721. METHODS： The human hepatoma cells T7721 which was stably transfected with HAb18G/CD147 and non-transfected 7721 were cultured. The total RNA extracted from T7721 and 7721 was dye-swap-labeled with fluorescein （Cy5 and Cy3 ） and then analyzed using signal transduction-related microarray. RT-PCR was employed to investigate the differential expression of the pivotal molecule （ βig-h3 ） primarily screened by microarray in T7721 and 7721. RESULTS： ① By differential analysis of microarray, we found that the mRNA expression of TGFβ-induced molecule βig-h3 in T7721 was upregulated significantly, as 4.4 times as that in 7721. Whereas the expression of TGFβ1 mRNA hadn＇t obvious difference between the two cell lines. ② The results of RT-PCR suggested that the expression of βig-h3 mRNA in T7721 was much higher than that in 7721 （ t = 14.42, P 〈 0.01 ）. CONCLUSION： The expression of TGFβ-induced molecule βig-h3 is positively correlated with HAb18G/CD147, and the overexpression of HAb18G/CD147 promotes the expression of βig- h3, which provides a sound basis for further investigating the role of HAb18G/CD147 in invasion and metastasis of human hepatoma cells.