环氧化酶2抑制剂对宫颈癌Hela细胞中HPV18-E6和COX-2表达的影响

The influence of COX-2 inhibitor on the expression of HPV18-E6 and COX-2 in Hela cells

目的:探讨环氧化酶2抑制剂Celecoxib对宫颈癌细胞Hela中HPV18-E6、COX-2表达及其细胞增殖和凋亡的影响。方法:用不同浓度Celecoxib处理Hela细胞株后,免疫细胞化学SP法检测HPV18-E6和COX-2蛋白的表达变化。RT-PCR检测HPV18-E6及COX-2 mRNA表达的改变。MTT法检测其对细胞增殖的影响。流式细胞仪测定细胞凋亡情况。Hoechst33258染色观察细胞的形态学变化。结果:Celecoxib作用Hela细胞后HPV18-E6和COX-2基因mRNA及蛋白表达量明显低于对照组,且随着药物浓度增加表达呈梯度下降。各组之间差异具有显著性(P<0.05),且两者之间具有显著相关性(P<0.01)。Celecoxib抑制Hela细胞增殖,作用呈量-效关系(P<0.05)。流式细胞仪显示各实验组凋亡率随浓度升高而增加,与对照组之间有显著差异(P<0.05)。Hoechst33258荧光染色显示经20μmol/L药物作用Hela细胞24h后表现为典型调亡小体。结论:COX-2抑制剂Celecoxib可以同时下调宫颈癌Hela细胞中HPV18-E6和COX-2表达、抑制Hela细胞的增殖、促进其凋亡。

Objective：To explore the influence of COX - 2 inhibitor Celecoxib on the expression of HPV18 - E6 and COX - 2 in Hela cells and its viability and apoptosis. Methods：Human uterine cervix cancer Hela cells were treated with different concentrations of Celecoxib. The expression of HPV18 - E6 and COX - 2 were detected by im-munocytochemical SP method and reverse transcription - polymerase chain reaction. Cell survival viability was tested by using the MTT assay. The change of cell apoptosis was detected by flow cytometry and its morphology was observed under microscope after Hoechst staining. Results：The expression of HPV18 - E6 and COX - 2 protein was inhibited by COX - 2 inhibitor,with time/ dose - dependent manner and they were positively related in Hela cells. The growth was inhibited and the apoptosis of Hela cells was induced by COX - 2 inhibitor. Conclusion：COX - 2 inhibitor can lower HPV18 - E6 and COX - 2 expression at the same time inhibit Hela cells viability and enhance its apoptosis.