siRNA干扰MeCP2基因对宫颈癌Hela细胞的生物学影响

Biological effects of siRNA targeting MeCP2 gene on cervical cancer Hela cells

目的研究甲基化Cp G结合蛋白2(Me CP2)对宫颈癌Hela细胞的生物学影响以及其分子机制。方法应用si RNA干扰Me CP2表达;MTT比色法分析Me CP2对宫颈癌Hela细胞增殖活力的影响;采用流式细胞仪分析对细胞周期和细胞凋亡的影响;Western blotting观察Me CP2 si RNA对Cyclin D1、P-ERK1/2和P-p38表达的影响。结果 Me CP2 si RNA组与阴性对照组相比,Me CP2在m RNA和蛋白水平的表达均显著下调;Me CP2 si RNA抑制宫颈癌Hela细胞增殖;S和G2/M期细胞数量显著减少,G1/G0期细胞数量显著增加;Me CP2 si RNA组早期凋亡和晚期凋亡显著增加;Cyclin D1和P-ERK1/2表达显著下调;P-p38表达显著上调,均P<0.01。结论 Me CP2通过调控P-ERK1/2和Cyclin D1的表达促进宫颈癌Hela细胞增殖,经过抑制p38 MAPK信号通路抑制了细胞凋亡。

Objective To investigate the biological effects of methyl-Cp G-binding protein 2（MeCP2） on cervical cancer Hela cells and its molecular mechanism. Methods The si RNA interference was used to knockdown MeCP2 expression in Hela cells. MTT assay was used to analyze the effects of MeCP2 on the proliferation of cervical cancer Hela cells. The effects of MeCP2 on the cell cycle and apoptosis of cervical cancer Hela cells was observed by flow cytometer. The effects of MeCP2 si RNA on the expression of Cyclin D1, P-ERK1/2and P-p38 were analyzed by Western blotting. Results MeCP2 expression at the levels of m RNA and protein significantly decreased in MeCP2 si RNA group compared to negative control group（P 〈 0.01）. MeCP2 si RNA inhibited the proliferation of cervical cancer Hela cells. The number of S and G2/M phase cells significantly was decreased in MeCP2 si RNA group compared to negative control group（P 〈 0.01）, meanwhile the number of G1/G0 phase cells remarkably was increased（P 〈 0.01）. The number of early apoptotic and lateapoptotic cells was remarkably increased in MeCP2 si RNA group compared to negative control group（P 〈 0.01）. It was found that Cyclin D1 and P-ERK1/2 proteins significantly were decreased in MeCP2 si RNA group compared to negative control group（P 〈 0.01）, P-p38 proteins remarkably increased（P 〈 0.01）. ConclusionMeCP2 promotes proliferation of cervical cancer Hela cells through regulating P-ERK1/2 and Cyclin D1 expression, and inhibits apoptosis by suppressing P-p38 MAPK signal pathway.