发育相关转录因子GATA4在小鼠肝癌细胞中的表达及其意义  

Expression of GATA4 in mice hepatocarcinoma cells and its significance

作　　者：任传路[1] 刘善荣[1] 杨玲[1] 王凤玫[1] 熊俊[1] 汤淑萍[1] 刘厚奇[1] 

机构地区：[1]第二军医大学基础医学部组织胚胎学教研室,上海200433

出　　处：《第二军医大学学报》2005年第4期361-365,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(90208026).

摘　　要：目的:探讨转录因子GATA4在肝癌细胞中的表达及其在肝癌发生中的作用.方法:将GATA4真核表达载体转染小鼠正常肝细胞,RT-PCR、免疫组化鉴定转染效果,G418筛选稳定表达细胞克隆;进行细胞计数并绘制未转染、空转染、转染GATA4组及肝癌细胞组细胞生长曲线;RT-PCR法比较成纤维细胞(3T3,L1)、GATA4转染肝细胞(BNLG,L2)、未转染肝细胞(BNL,L3)、肝癌细胞株H22细胞(H22,L4)、肝癌细胞株Hepa1-6细胞(Hepa1-6,L5)和小鼠孕13.5 d胎肝细胞(孕13.5d胎肝,L6)的GATA4、胎蛋白转录因子(FTF)和甲胎蛋白(AFP)mRNA的表达.结果:GATA4真核表达载体转染效果较好;小鼠肝癌细胞株高表达GATA4 mRNA而正常肝细胞几乎不表达;GATA4转染组小鼠肝细胞增殖速度明显大干未转染和空转染组,但要小于肝癌细胞组.L1~L6的GATA4、FTF、AFP mRNA表达强度分别为0.04±0.02,0.28±0.03,0.04±0.02,0.26±0.03,0.32±0.04,0.24±0.03;0.04±0.02,0.26±0.02,0.08±0.02,0.22±0.02,0.26±0.02,0.26±0.01;0.04±0.01,0.37±0.03,0.14±0.02,0.50±0.03,0.51±0.03,0.49±0.03.L1、L3与L2、L4、L5、L6比较差异显著(P<0.01),说明正常肝细胞的GATA4、FTF、AFP mRNA表达明显低于肝癌和胎肝细胞,转染后表达增高.结论:GATA4在肝癌细胞中高表达,并能够促进正常肝细胞增殖,促进FTF、AFP mRNA的表达,可能参与肝癌的发生、发展.Objective:To study the expression of transcription factor GATA4 in hepatocarcinoma cells and its effect on hepatocarcinogenesis. Methods: After normal liver cell line BNL was transfected with recombinant plasmid pcDNA3.1-GATA4, stably expressed cell clones were selected with G418 and further identified by immunohistochemistry and RT-PCR. Then cell growth curves were produced from the data of wild type BNL cells, mock-transfected cells, pcDNA3.1-GATA4 transfected cells (BNLG) and control Hepa1-6 cells. The expressions of GATA4 mRNA, FTF mRNA and AFP mRNA in 3T3 (L 1), BNLG (L 2), BNL (L 3), Hepa1-6 (L 4), H22 (L 5) and fetal liver tissue (13.5 dpc, L 6) were compared by RT-PCR. Results: The transfection proved to be effective. Over expression of GATA4 mRNA was detected in the hepatocarcinoma cells and little expression in normal ones. The proliferation of BNLG was much more rapid than that of BNL and mock-transfected cells, and slower than that of Hepa1-6. The expression intensities of GATA, FTF, AFP mRNA from L 1 to L 6 were 0.04±0.02,0.28± 0.03,0.04± 0.02,0.26±0.03,0.32±0.04,0.24±0.03; 0.04±0.02,0.26±0.02,0.08±0.02,0.22±0.02,0.26±0.02, 0.26±0.01; 0.04±0.01,0.37±0.03,0.14±0.02,0.50±0.03,0.51±0.03,0.49±0.03, respectively. There were significant differences between L 1,L 3 and L 2,L 4,L 5,L 6 (P<0.01),implying that the expressions of GATA, FTF, and AFP mRNA in normal liver cell lines were much lower than that in hepatocarcinoma cells and fetal liver cells, and became higher after transfection. Conclusion: Overexpression GATA4 in hepatocarcinoma cells not only promotes the proliferation of normal liver cells but also up-regulates the expression of FTF and AFP mRNA, which may be implied in the development and progression of hepatocarcinoma.

关 键 词：肝肿瘤 GATA4 肝癌细胞 甲胎蛋白 

分 类 号：R735.7[医药卫生—肿瘤]