中文摘要：

目的 探讨PNPLA3基因I148M多态性对人肝癌细胞株Huh-7细胞周期的影响及其相关机制. 方法 利用构建成功的稳定表达PNPLA3基因野生型和PNPLA3基因I148M突变型的Huh-7细胞株,以空载体细胞为对照,流式细胞术检测转染的Huh-7细胞周期,Western blot方法检测细胞周期调节因子Cyclin D1、p53的蛋白表达情况.荧光定量PCR检测Cyclin D1、p53的mRNA表达情况.两组间比较用独立样本t检验. 结果 三组间各细胞周期细胞所占比例的两两比较显示野生型组细胞周期G1期（58.53％±0.35％）,G2/M期（24.87％±0.60％）,S期（16.60％±0.26％）参数与对照组对应各期（G1期59.27％±0.15％;G2/M期24.23％±0.31％;S期16.50％±0.26％）之间差异无统计学意义（P＞ 0.05）;突变型组与野生型组相比,G1期细胞比例减少（突变型G1期38.37％±0.21％,P＜0.05）,S期与G2/M期细胞比例相应增加（突变型S期27.47％±0.35％,G2/M期34.17％±0.15％,P＜0.05）.与对照组相比,突变型组的p53 mRNA相对表达量明显上调（对照组1.06±0.41,突变型组6.54±0.34;P＜0.05）;野生型组p53 mRNA相对表达量差异无统计学意义（1.66±0.30,P＞0.05）.Cyclin D1 mRNA各组中的相对表达量差异均无统计学意义（对照组1.00±0.10,野生型组1.06±0.03,突变型1.11±0.04;P＞0.05）.结论 PNPLA3基因I148M多态性通过上调p53的表达引起Huh-7细胞周期紊乱.

英文摘要：

Objective To investigate the cell cycle of Huh-7 cells affected by I148M polymorphism of PNPLA3 gene and the possible mechanisms.Methods Huh-7 cells which could respectively overexpress PNPLA3 wild type and I148M variant were cultured and Huh-7 cells with zero load plasmids were used as matched control,Flow cytometry was conducted to detect the cell cycles of these 3 type of Huh-7 cells and western blot and realtime fluores-cence quantitative PCR were applied to investigate the expression of regulatory factors （Cyclin D1 and p53） of cell cycle.t-test was used in statistical analysis.Results Cell cycle phase distribution was presented by the proportion of cells in each phases （%）,compared with the control group,the cell cycle phase distribution （G1 phase 59.27±0.15,G2/M phase 24.23±0.31,S phases 16.50±0.26） had no differences in wild type group （G1 phase 58.53±0.35,G2/M phase 24.87±0.60,S phases 16.60±0.26;P〈0.05）.While between variant type group and wild type group,G1 phase was significantly decreased （variant type group G phase38.37±0.21,P〈 0.05）,S phase and G2/M phase were increased （variant type group S phase 27.47±0.35,P 〈 0.05;G2/M phase 34.17±0.15,P 〈 0.05）,respectively.compared with control group,the relative expression of P53 mRNA in variant type group was significantly upregulated （control group 1.06±0.41,variant type group 6.54±0.34;P〈0.05） and there was no statistical significance in wild type group （1.66±0.30,P 〉 0.05）;Cyclin D1 expression showed no statistical si gnificance in any of these three groups,control group 1.00±0.10,wild type group 1.06±0.03,variant type group,1.11±0.04;P〉 0.05）.Conclusion I148M polymorphism of PNPLA3 gene affects cell cycles of Huh-7 cells via up-regulatating P53.