GRIM-19对子宫颈癌细胞株HeLa细胞Survivin基因表达的影响

目的:探讨稳定转染GRIM-19对子宫颈癌细胞株HeLa细胞凋亡抑制基因Survivin的表达以及细胞增殖的影响。方法:采用脂质体转染法将GRIM-19重组质粒转染子宫颈癌细胞株HeLa细胞,建立稳定表达GRIM-19的HeLa细胞株,采用RT-PCR检测HeLa细胞转染GRIM-19后mRNA水平表达情况,采用WesternBlot检测稳定转染后GRIM-19与Survivin的表达情况,应用细胞计数法观察各组细胞增殖水平。结果:建立稳定的GRIM-19基因表达的HeLa细胞株,命名为HeLa/G19细胞,对照组细胞命名为HeLa/Con细胞。RT-PCR检测显示GRIM-19mRNA表达明显增加,WesternBlot检测显示GRIM-19的蛋白表达显著升高、Survivin蛋白的表达水平显著降低。细胞计数法实验显示HeLa/GRIM-19细胞增殖较HeLa/Con细胞明显减慢(P<0.05)。结论:GRIM-19可以抑制子宫颈癌细胞Survivin的表达,阻滞细胞的生长,可能是子宫颈癌治疗的新靶点。

Effect of GRIM-19 on expression of Survivin gene in cervical cancer cell line HeLa cells

Objective:To explore the effects of GRIM-19 on expression of Survivin gene in cervical cancer cell line HeLa cells and cell proliferation.Methods:The cancer cell line HeLa cells were transferred with GRIM-19 recombinant plasmid by liposome infection protocol,HeLa cells expressing GRIM-19 stably were constructed,RT-PCR was used to detect the expression level of GRIM-19 mRNA after transfection,Western blot was used to detect the expression levels of GRIM-19 and Survivin after transfection,cytometry was used to observe the proliferative levels of cells in different groups.Results:Stable HeLa cell line expressing GRIM-19 gene was established and named as HeLa/G19 cells,for control group,the cell line was named as HeLa/Con cells;the results of RT-PCR indicated that the expression of GRIM-19 mRNA increased significantly,the results of Western blot indicated that the expression of GRIM-19 protein increased significantly,while the expression of Survivin protein decreased significantly;the results of cytometry indicated that the proliferation of HeLa/GRIM-19 cells was slower than that of HeLa/Con cells (P < 0.05).Conclusion:GRIM-19 can inhibit the expression of Survivin in cervical cancer cells and block cell proliferation,which may be a new target for treatment of cervical cancer.