佛波酯对HaCaT细胞环氧合酶-2表达水平的影响

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0.1，1.0，10 mg/L佛波酯处理24 h后COX-2 mRNA和蛋白表达情况。结果 对照组HaCaT 细胞COX-2 mRNA和蛋白微弱表达或不表达，1.0，10.0 mg/L佛波酯组COX-2 mRNA和蛋白表达水平均显著高于对照组和0.1 mg/L佛波酯组（P值均 < 0.01），而且10.0 mg/L佛波酯组均高于1.0 mg/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。

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

Objective To investigate the effect of phorbol-12-myristate-13-acetate(PMA)on cyclooxygenase-2 (COX-2) mRNA and protein expression in cultured human HaCaT keratinocytes,and the mechanism for cytotoxity of PMA against keratinocytes.Methods RT-PCR and Westem blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT ells 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 df COX-2 mRNA and protein in HaCaT cells;incubation witll 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 difrerence was statistically significant (all P＜0.01).Conclusion These results suggest that PMA may be involved in keratinocyte tumorigenesis by upregulating he expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.