不同蛋白激酶C同工酶亚型对卵巢癌细胞耐药性的影响

The effect of protein kinase C isoforms on the drug-resistance of ovarian cancer cell lines

目的 :检测不同的蛋白激酶 (PKC)同工酶亚型在人卵巢癌耐药细胞中的表达和功能 ,探讨它们与P 糖蛋白 (P gp)介导的多药耐药性之间的关系。方法 :采用免疫荧光及免迹印迹方法检测PKCα和PKCβI在人卵巢A2 780细胞和A2 780 /ADM耐药细胞中的表达 ,免疫印迹法检测P gp的表达 ,并用流式细胞仪测定细胞内柔红霉素 (DNR)及罗丹明 1 2 3 (Rh1 2 3 )的荧光强度。结果 :在A2 780 /ADM细胞中 ,PKC与P gp的表达较A2 780细胞增高 ,PKCβI表达在两者间无明显改变 ;在A2 780 /ADM中DNR及Rh1 2 3的荧光强度较A2 780中降低。A2 780 /ADM细胞与抗PKCα抗体共同卵孕育后细胞内DNR含量上升 ,与十字孢碱 (SP)作用后细胞内Rh1 2 3泵出减少 ,积聚量升高。结论 :PKCα可能在P

Objective:To explore the expression and function of protein kinase C isoforms in drug resistant human ovarian cancer cell,and to determine the relationship with the multidrug resistance(MDR) induced by P glycoprotein(P gp).Metheds:The expression of PKCα and PKCβI were monitored by immunofluorescence technique and Western blot in adriamycin resistant(A2780/ADM) and adriamycin sensitive(A2780) cell lines.Western blot was also used to determin P gp expression.The fluorescence of daunorubicin(DNR) and rhodamine 123(Rh123) in A2780/ADM and A2780 were detected by flow cytometry.Results:The expression of PKCα and P gp in A2780/ADM was higher than that in A2780,but there was no change with PKCβI in both A2780/ADM and A2780.The fluorescence of DNR and Rh123 in A2780/ADM was lower compared with A2780.An increased DNR uptake of A2780/ADM revealed when Co incubated with anti PKCα.The efflux of Rh123 inducd by P gp in A2780/ADM was reduced after treatd with staurosporine(SP).Conclusion:PKCα may play an important role in multidrug resistance of human ovarian cancer induced by P gp.