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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

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【作者】 王薇杨晓葵邬素芳高庆蕾李静陈刚李辅军王世宣卢运萍马丁

【Author】 Wang Wei,Yang Xiaokui,Wu Sufang,et al.Department of Obstetrics and Gynecology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030

【机构】 华中科技大学同济医学院附属同济医院妇产科华中科技大学同济医学院附属同济医院妇产科 武汉430030武汉430030武汉430030

【摘要】 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 gp介导的人卵巢癌细胞多药耐药中起重要作用

【Abstract】 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.

【基金】 国家重点基础研究发展项目 (2 0 02CB51 31 0 0 );国家杰出青年科学基金资助项目(30 0 2 50 1 7)
  • 【文献出处】 现代妇产科进展 ,Current Advances In Obstetrics and Gynecology , 编辑部邮箱 ,2003年05期
  • 【分类号】R737.31
  • 【被引频次】6
  • 【下载频次】74
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