蛋白激酶C在人骨肉瘤细胞多药耐药中的作用及机制  

Effect of protein kinase C on multidrug resistance in human osteosarcoma cells and its underlying mechanism

作　　者：陈渝[1] 邓忠良[2] 

机构地区：[1]重庆市第九人民医院骨科,重庆400700 [2]重庆医科大学附属第二医院骨科,重庆400010

出　　处：《第三军医大学学报》2011年第18期1900-1903,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30772211)~~

摘　　要：目的研究蛋白激酶C(PKC)主要亚型在人骨肉瘤143B细胞中的表达情况及其与143B细胞耐药的关系。方法采用Western blot法检测PKC主要亚型α、β和γ磷酸化形式p-PKCα、p-PKCβ、p-PKCγ和多药耐药蛋白-1(mutidrugresistance protein-1,MDR-1)在人骨肉瘤143B/WT野生型细胞和143B/ADM耐药细胞中的表达水平,通过PKC特异性抑制剂staurosporine抑制PKC对MDR-1蛋白表达的影响;CCK法检测药物敏感性;荧光分光光度计法检测细胞内阿霉素浓度,流式细胞术检测细胞凋亡水平和细胞外排罗丹明水平。结果相对于143B/WT野生型细胞,143B/ADM耐药细胞中p-PKCα表达显著增加,而p-PKCβ和p-PKCγ表达则无明显差异,同时143B/ADM耐药细胞中MDR-1表达也明显上调;143B/ADM耐药细胞对阿霉素的耐药指数为15.7,而加入staurosporine抑制143B/ADM细胞PKC表达后,细胞耐药指数下降为5.5(P<0.01);143B/ADM耐药细胞接受ADM处理后,细胞凋亡率为(28.41±4.33)%,而其PKC表达被抑制后,细胞内阿霉素浓度增加的同时(P<0.01),细胞凋亡率增加为(70.63±4.80)%(P<0.01),外排罗丹明减少,同时MDR-1蛋白表达明显下调。结论人骨肉瘤耐药细胞中过表达的PKCα可通过调控MDR-1参与骨肉瘤耐药。Objective To study the expression of major protein kinase C（PKC） subtypes in human osteosarcoma 143B cells and its relation with drug resistance to 143B cells.Methods Expression levels of p-PKCα,p-PKCβ,p-PKCγ and multidrug resistance protein 1（MDR-1） in wild human osteosarcoma cells or 143B/ADM-drug resistant cells were measured by Western blotting.Effect of PKC on expression of MDR-1 was inhibited by the PKC special inhibitor,staurosporine.Drug sensitivity was measured by CCK assay.Concentration of adriamycin（ADM） in 143B cells was measured by fluorescence spectrometry.Cell apoptosis and extracellular rho123 levels were measured by flow cytometry.Results The expression level of p-PKCα was significantly higher in 143B/ADM-drug resistant cells than in 143B/WT cells.However,no significant difference was found in expression of p-PKCβ and p-PKCγ.The expression of MDR-1 in 143B/ADM-drug resistant cells was significantly up-regulated.The drug-resistant index to ADM was 15.7 in 143B/ADM-drug resistant cells but decreased to 5.5 after the expression of PKC in 143B/ADM cells was inhibited by staurosporine（P0.01）.The apoptosis rate of 143B/ADM-drug resistant cells was 28.41%±4.33% and increased to 70.63%±4.80%,while the extracellular rho123 level and MDR-1 expression level were significantly down-regulated after the expression of PKC in 143B/ADM cells was inhibited by staurosporine（P0.01）.Conclusion Over-expression of PKCα in drug-resistant human osteosarcoma 143B cells may attenuate drug resistance by regulating MDR-1 expression.

关 键 词：人骨肉瘤143B细胞 阿霉素 蛋白激酶C 多药耐药蛋白-1 

分 类 号：R341.5[医药卫生—基础医学] R738.1