GRIM-19表达对子宫颈癌HeLa细胞增殖的影响

Effects of expression of GRIM-19 on proliferation of cervical cancer cell line HeLa

目的:通过体内和体外实验探讨GRIM-19表达对子宫颈癌HeLa细胞增殖的影响。方法:采用脂质体转染法将GRIM-19重组质粒转染HeLa细胞,建立稳定表达GRIM-19的细胞株;采用RT-PCR和Western印迹法检测HeLa细胞转染GRIM19重组质粒前后GRIM-19 mRNA和蛋白水平的表达情况;应用MTT法检测各组细胞增殖水平,并将HeLa/con、HeLa/GRIM-19细胞接种到裸鼠皮下以检测荷瘤情况;同时对移植瘤检测其GRIM-19 mRNA及其下游STAT3 mRNA的表达水平。结果:建立稳定表达GRIM-19基因的HeLa/GRIM-19细胞株。RT-PCR结果显示GRIM-19基因在HeLa/GRIM-19细胞中mRHA表达明显增加,Western印迹法检测同样发现GRIM-19蛋白的表达显著升高,而STAT3的表达明显受到抑制。MTT检测结果显示HeLa/GRIM-19细胞增殖较HeLa/con明显减慢(P<0.05),其接种后肿瘤生长速度亦明显减慢(P<0.01)。移植瘤的RT-PCR结果表明GRIM-19表达升高、STAT3表达降低。结论:GRIM-19的表达可以抑制子宫颈癌细胞的增殖,可能作为子宫颈癌治疗的一个新靶点。

Objective：To explore the effects of expression of GRIM-19 on the proliferation of cervical cancer HeLa cells in vivo and in vitro. Methods：The stable transfected cervical cancer cell line HeLa/GRIM-19 was established by transfecting recombinant pGRIM-19 into HeLa cells via liposome mediation. The mRNA and protein expressions in HeLa cells stably transfected with GRIM-19 were investigated by RT-PCR and Western blotting before and after transfection. Cell proliferation was assessed by MTT assay. HeLa/con and HeLa/GRIM-19 were subcutaneously inoculated into nude mice and the growth of xenografted tumor was observed. The mRNA expressions of GRIM-19 and STAT3 were determined with RT-PCR. Results：HeLa cells with stable transfection of GRIM-19 were established. RT-PCR showed that the expression of GRIM-19 mRNA was significantly enhanced and Western blotting demonstrated that GRIM-19 protein expression was markedly increased,and the expression of STAT3 protein was greatly inhibited in HeLa/GRIM-19 cells. MTT assay showed that compared with HeLa/con cells,the speed of proliferation of HeLa/GRIM-19 cells was slowed down （P〈0.05）. The growth of implanted tumor of HeLa/GRIM-19 cells was significantly slowed down （P〈0.01）.The expression of GRIM-19 mRNA was markedly increased,while STAT3 mRNA was decreased in implanted tumors. Conclusion：The expression of GRIM-19 inhibits the proliferation of cervical cancer cells,suggesting that GRIM-19 may be a novel target for the treatment of cervical cancer.