FOXP3过表达对HPV感染阳性宫颈癌细胞增殖和凋亡的影响

摘要

目的:探讨叉状头/翅膀状螺旋转录因子(Foxp3)过表达对人乳头状瘤病毒(HPV)感染阳性宫颈癌细胞增殖、凋亡及细胞免疫的影响.方法:利用脂质体介导转染方法,建立Foxp3过表达HPV感染阳性SiHa细胞株,作为Foxp3过表达阳性细胞,同时利用pcDNA3.1空载体转染SiHa细胞株,作为本研究阴性对照,同时设置空白对照组,采用实时荧光定量聚合酶链式反应(qRT-PCR)方法及Western blot法检测三组Foxp3 mRNA及蛋白表达水平,采用酶联免疫吸附试验(ELISA)检测SiHa细胞中白细胞介素8(IL-8)、白细胞介素10(IL-10)水平,采用四甲基偶氮唑蓝(MTT)实验检测SiHa细胞增殖能力,采用利用流式细胞仪(FCM)检测SiHa细胞周期及凋亡率情况.结果:过表达组Foxp3 mRNA及蛋白表达水平明显高于阴性对照组及空白对照组,差异有统计学意义(P<0.05),阴性对照组及空白对照组Foxp3 mRNA及蛋白表达水平比较,差异无统计学意义(P>0.05);与阴性对照组及空白对照组比较,各时间点过表达组IL-8水平明显降低,36 h、48 h IL-10水平明显升高;MTT结果显示,Foxp3过表达可明显促进SiHa细胞增殖,FCM检测显示,Foxp3过表达可明显抑制SiHa细胞凋亡,抑制G0/G1期阻滞.结论:Foxp3过表达可促进HPV感染阳性宫颈癌细胞增殖,抑制细胞凋亡.%Objective:To explore the influence of forkhead/winged helix transcription factor(FOXP3)overexpres-sion on cell proliferation and apoptosis of human papilloma virus(HPV)infected positive cervical cancer cells.Meth-ods:Foxp3 overexpression of HPV infected positive SiHa cell lines were developed by using liposome mediated trans-fection method,as Foxp3 overexpression positive cell,and transfected SiHa cell lines by using pcDNA3.1 empty carri-er were as the negative control group in the study,and meanwhile,the blank control group were set.Foxp3 mRNA and protein expression levels were detected in the three groups by using real-time fluorescence quantitative polymer-ase chain reaction(qRT-PCR)and Western blot method.Interleukin 8(IL-8)and interleukin 10(IL-10)levels in SiHa cells were tested by using Enzyme linked immunosorbent assay(ELISA)method. SiHa cell proliferation ability were tested by using methylthiazolyldiphenyl-tetrazolium bromide(MTT)experiment,and SiHa cell cycle and apoptosis rate were detected by using flow cytometry(FCM).Results:The expression levels of Foxp3,mRNA and pro-tein in the overexpression group were significantly higher than those in the negative control group and the blank control group,with statistically significant differences(P<0.05).There were no significant differences in the expression lev-els between the negative control group and the blank control group(P>0.05).Compared with those in the negative control group and the blank control group,IL-8 level decreased significantly in different time points,and IL-10 lev-el increased significantly at 36 h and 48 h in the overexpression group.MTT result showed that Foxp3 overexpression could significantly promote the proliferation of SiHa cells.FCM showed that Foxp3 overexpression could significantly inhibit the apoptosis of SiHa cells,and inhibit G0/G1blocking.Conclusion:FOXP3 overexpression can promote the proliferation of HPV infected positive cervical cancer cells,and inhibit the apoptosis.