Pin1通过调控脂质代谢关键酶影响宫颈癌细胞的增殖及凋亡

Effects of Pin1 on the Proliferation and Apoptosis of Cervical Cancer SiHa Cells by Regulating Key Enzymes of Lipid Metabolism

目的 探讨下调Pin1表达对宫颈癌SiHa细胞的脂质合成及增殖、凋亡的影响。方法 Western Blot检测Pin1在Hela、SiHa和H8细胞的表达情况;慢病毒转染技术构建Pin1低表达稳转细胞株,根据不同处理分为对照组(shPin1-NON)、敲低组1(shPin1-1)和敲低组2(shPin1-2),GEPIA2分析脂质代谢关键酶(ACC1、FASN)在宫颈癌及非肿瘤组织中的表达;Western Blot和qRT-PCR检测ACC1、FASN蛋白和mRNA的表达;Western Blot检测Caspase-8、BCL-2、C-myc蛋白表达;CCK-8检测细胞增殖能力;克隆形成实验检测细胞集落形成能力;流式细胞术检测细胞凋亡情况。结果 Pin1在宫颈癌细胞中高表达,且在SiHa细胞中表达最高(P <0.05);Pin1低表达慢病毒有效下调了Pin1表达水平;宫颈癌中ACC1、FASN mRNA表达高于非肿瘤组织;shPin1-1组和shPin1-2组ACC1、FASN蛋白和mRNA表达均低于shPin1-NON组(P <0.05);shPin1-1组和shPin1-2组相较于shPin1-NON组Caspase-8蛋白表达被上调,而BCL-2、C-myc的表达均被抑制;shPin1-1组和shPin1-2组细胞增殖能力低于shPin1-NON组(P <0.05);shPin1-1组和shPin1-2组细胞集落形成能力小于shPin1-NON组(P <0.05)。shPin1-1组和shPin1-2组细胞总凋亡率相较shPin1-NON组显著增加(P <0.05)。结论 Pin1低表达有效下调宫颈癌SiHa细胞脂质代谢关键酶,抑制细胞增殖,诱发细胞凋亡,为靶向治疗宫颈癌提供新思路。

Objective To investigate the effect of down-regulation of Pin1 expression on lipid synthesis,proliferation and apoptosis of cervical cancer SiHa cells.Methods Western Blot was used to detect the expression of Pin1 in Hela,SiHa and H8 cells;Lentiviral transfection technology constructed Pin1 low-expression stable transduction cell line,which was divided into the control group(shPin1-NON),knockdown group1(shPin1-1)and knockdown group2(shPin1-2) according to different treatments;GEPIA2 was used to analyze the expression of key enzymes of lipid metabolism(ACC1,FASN) in cervical cancer and non-tumor tissues and the expression of ACC1 and FASN in cervical cancer and non-tumor tissues;Western Blot and qRT-PCR were used to detect the ACC1,FASN protein and mRNA expression;Western Blot was used to detect the Caspase-8,BCL-2,C-myc protein expression;CCK-8 was used to detect the cell proliferation;Clone formation assay was used to detect the cell colony formation ability and flow cytometry was used to detect the cell apoptosis.Results Pin1 was highly expressed in cervical cancer cells,and the highest expression was in Si Ha cells;Pin1 low expression lentivirus effectively down-regulated the Pin1 expression level;ACC1 and FASN m RNA expressions in cervical cancer were higher than those in non-tumor tissues;The protein and m RNA expressions of ACC1 and FASN in sh Pin1-1 group and sh Pin1-2 group were lower than those in the sh Pin1-NON group;the sh Pin1-1 and sh Pin1-2 groups were upregulated compared with the sh Pin1-NON group,while the expression of BCL-2 and C-myc was up-regulated.The expression was inhibited;The proliferation ability of sh Pin1-1 group and sh Pin1-2 group was lower than that of sh Pin1-NON group;The colony formation ability of sh Pin1-1 group and sh Pin1-2 group was lower than that of sh Pin1-NON group.Compared with the sh Pin1-NON group,the total apoptotic rate of cells in the sh Pin1-1 and sh Pin1-2 groups was significantly increased.(P < 0.05).Conclusion Low expression of Pin1 can effectively downregulate fatty acid synthase in cervical cancer Si Ha cells,inhibit cell proliferation,and induce cell apoptosis,which provides a new idea for targeted therapy of cervical cancer.