佛波酯对HaCaT细胞环氧合酶-2表达水平的影响

Effect of phorbol-12-myristate-13-acetate on cyclooxygenase-2 expression in human HaCaT kerafinoeytes

目的研究佛波酯对培养的人永生化角质形成细胞株HaCaT细胞环氧合酶-2（COX-2）表达水平的影响，探讨佛波酯的皮肤细胞毒性机制。方法以体外培养的HaCaT细胞为对象，采用RT-PCR和Western印迹方法，检测HaCaT细胞经0．1，1．0，10mg／L佛波酯处理24h后COX-2mRNA和蛋白表达情况。结果对照组HaCaT细胞COX-2mRNA和蛋白微弱表达或不表达，1．0，10．0mg／L佛波酯组COX-2mRNA和蛋白表达水平均显著高于对照组和0．1mg／L佛波酯组（P值均〈0．01），而且10．0mg／L佛波酯组均高于1．0mg／L佛波酯组，差异均有统计学意义（P〈0．01）。结论佛波酯可能通过上调HaCaT细胞表达COX-2，促进前列腺素的合成释放，参与皮肤角质形成细胞恶性肿瘤的发生。

Objective To investigate the effect ofphorbol-12-myristate-13-acetate （PMA） on cyclooxygenase-2 （COX-2） mRNA and protein expression in cultured human HaCaT kemtinocytes, and the mechanism for cytotoxity of PMA against keratinocytes. Methods RT-PCR and Western blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT cells at 24 hours after the treatment with various concentrations of PMA （0.1, 1.0, 10 mg/L）. Results Without any treatment, there was no or a weak expression of COX-2 mRNA and protein in HaCaT cells; incubation with PMA resulted in the induction of the expression of COX-2 in HaCaT cells. The expression levels of COX-2 mRNA and protein in 10 mg/L PMA-pretreated HaCaT cells were significantly higher than those in 1.0 mg/L PMA-pretreated HaCaT cells, which was in turn higher than that in 0.1 mg/L PMA-pretreated cells and untreated cells; the difference was statistically significant （all P 〈 0.01 ）. Condusion These results suggest that PMA may be involved in kemtinocyte tumorigenesis by upregulating the expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.