表没食子儿茶素没食子酸酯对HPV11早期基因E6、E7mRNA表达的影响

目的 利用人工构建的含人乳头瘤病毒(HPV)11型基因组的HaCaT细胞(简称为HPV11.HaCaT细胞),研究表没食子儿茶素没食子酸酯(EGCG)对该细胞生长的抑制作用,以及受试物对HPV11早期基因E6和E7 mRNA表达的影响。方法 在HaCaT及HPV11.HaCaT细胞培养基中添加不同浓度的EGCG后,应用MTT比色法检测受试物对细胞生长增殖的影响;用相对荧光定量PCR法检测HPV11.HaCaT细胞中HPV11 E6、E7 mRNA表达的改变。结果 EGCG能抑制HaCaT和HPV11.HaCaT细胞的生长,不同浓度EGCG作用下,细胞生存率为52%~95%、64%~90%,呈浓度和时间依赖趋势。EGCG作用于细胞12h和24h后,HPV11.HaCaT细胞中HPV11 E6、E7基因的mRNA表达水平明显下降(P<0.05)。结论 EGCG在体外能抑制含HPV11基因组的HaCaT细胞生长,并能抑制HPV11功能基因E6、E7表达。

Effect of (-)-epigallocatechin-3-gallate on Expression of HPV11 E6 and E7 mRNA

Objective Using of the HaCaT cells which contain the whole genes of human papillomavirus type 11 (HPV11.HaCaT), to study the effect of (-)-epigallocatechin-3-gallate (EGCG) on cell proliferation and expression of HPV11 E6 and E7 mRNA. Methods The HaCaT and HPV11.HaCaT cells were cultured and treated with different concentration of EGCG for different periods of time. The effects on proliferation of HaCaT and HPV11.HaCaT cells were examined by MTT assay. The alterations of HPV11 E6 and E7 mRNA level were examined by real-time fluorescence quantitative PCR. Results EGCG can inhibit the proliferation of HaCaT and HPV11.HaCaT in a dose- and time-dependent. EGCG in different concentration resulted in the cell survival rate ranging from 52% to 95% and 64% to 90%. After incubating the HPV11.HaCaT cells with EGCG for 12h and 24h, the expression of HPV11 E6 and E7 mRNA is inhibited obviously (P<0.05). Conclusion EGCG can inhibit the proliferation of HPV11.HaCaT cells and the expression of HPV11 E6 and E7 mRNA in vitro.