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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。
OBJECTIVE: To investigate the effect of GRIM-19 on the expression of Survivin and the proliferation of HeLa cells. METHODS: HeLa cells were transfected with GRIM-19 recombinant plasmid by lipofection method. HeLa cell line stably expressing GRIM-19 was established. The mRNA expression of GRIM-19 in HeLa cells was detected by RT-PCR The expression of GRIM-19 and Survivin in stable transfected cells was detected by Western Blot. The cell proliferation was observed by cell counting method. RESULTS: HeLa cell line with stable GRIM-19 gene expression was established and named as HeLa / G19 cells. The control group was named HeLa / Con cells. The expression of GRIM-19 mRNA was significantly increased by RT-PCR. Western Blot showed that the expression of GRIM-19 protein was significantly increased and the expression of Survivin protein was significantly decreased. Cell counting assay showed that the proliferation of HeLa / GRIM-19 cells was significantly 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 the growth of the cells, which may be a new target for the treatment of cervical cancer.