雌激素对人乳头状瘤病毒16型E6基因转染人永生化表皮细胞效率的影响

The effect of estrogen on human papilloma virus 16 E6 gene transfecting HaCat cell line

目的:探讨雌激素对人乳头状瘤病毒16(HPV 16)型E6基因转染人永生化表皮细胞效率的影响。方法:以不同浓度的17-β-雌二醇(E_2)处理人永生化表皮细胞系(Ha Cat)后,绘制其生长曲线,以细胞活性测定法(MTT)测定细胞活性,流式细胞仪检测细胞周期分布,溴脱氧尿嘧啶核苷法(Brd U)测定细胞周期时相。脂质体转染法将含HPV 16 E6基因的质粒转染入经不同药物处理后的细胞中,以Real-time PCR法测定细胞中该基因的表达。结果:浓度为10^(-7)~10^(-5) mol/L的E_2促进细胞数目的增加(P<0.05);浓度为10^(-8)~10^(-6) mol/L的E_2明显增强细胞活性(P<0.05);浓度为10^(-7)~10^(-5) mol/L的E_2使Ha Cat细胞的细胞周期时相缩短(P<0.05);不同浓度E_2处理组细胞周期的分布差异无统计学意义(P>0.05)。当Ha Cat细胞被HPV 16E6质粒转染,随着E_2浓度的增加,HPV 16 E6 m RNA表达明显增加(P<0.05)。结论:E_2可加速Ha Cat细胞生长与增殖,增加HPV 16 E6转染Ha Cat细胞的效率,E_2水平升高可能增加HPV感染的风险。

Objective： To determine the influence of 17-β-estradiol （E2） on human papilloma virus 16 E6 gene transfecting HaCat cell line and the possible mechanism. Methods： The HaCat cell line was incubated with E2 of varies concentration （10^-5, 10^-6, 10^-7, 10^-8, 10^-9, 0 mol/L） .The cell number was counted for 7 days to draw a growth curve. The cell viability was determined by MTT method. Flow cytometry was used to detect cell cycle distribution and BrdU to determine the time duration of each cell cycle phase. Then, the E2 treated cells was transfected by the plasmid containing HPV E6. The expression of E6 mRNA was determined by quantitative re- altime-PCR. Results： Compared with the control, at the concentration between 10^-7-10^-5 tool/L, E2 was observed to increase the number of HaCat cells, reduce the doubling time of cell number and increase the cell viability and E6 mRNA expression after transfection （P〈0.05）. Conclusion： E2 increased the cell growth and viability, en- hanced the txansfection efficiency of E6 on HaCat cells. The mechanism may be related to the acceleration of cell growth and cell viability.