KBV200细胞多药耐药与蛋白激酶C表达上调的关系(英文)  

Correlation of multidrug resistance with up-regulation of protein kinase C expression in KBV200 cells

作　　者：袁亚维[1] 孙爱民[1] 李传刚[2] 

机构地区：[1]南方医科大学南方医院放疗科,广东广州510515 [2]南方医科大学南方医院超声诊断科,广东广州510515

出　　处：《第一军医大学学报》2005年第11期1341-1343,1356,共4页Journal of First Military Medical University

基　　金：National Natural Science Foundation of China(39870809)

摘　　要：目的研究KBV200多药耐药细胞蛋白激酶C(PKC)的表达,探讨PKC的表达上调与肿瘤多药耐药的关系。方法KBV200细胞分对照组和佛波酯(PMA)组,PMA组应用200nmol/L的PMA预孵育细胞,而对照组不用。32P掺入法测定多药耐药KBV200细胞株的PKC活性,通过Westernblotting检测PKC各亚型表达和亚细胞分布。用MTT法检测细胞耐药性。结果PMA预孵育可提高KBV200细胞的PKC总活性和膜组分PKC活性,降低浆组分PKC活性(P<0.01)。PMA预孵育使膜组分PKCα表达增加,浆组分PKCα表达降低,膜组分PKCβ无明显变化,浆组分PKCβ的表达稍增强。PMA可升高长春新碱、阿霉素对KBV200细胞的IC50值(P<0.01)。结论PMA使KBV200细胞耐药性增加,可能与PKC表达上调有关。Objective To investigate protein kinase C （PKC） expression and its association with multidrug resistance （MDR） in KBV200 cells. Methods KBV200 cells were preincubated with PKC activator phorbol-12-myristate-13-acetate （PMA, 200 nmol/L） and PKC activity was assayed by measurement ofpeptide substrate ^32P incorporation from [γ-^32P]ATP, with the cells without PMA preincubation serving as the control. Western blotting was performed for assessing the expression ofPKC isoform, and the cell inhibition rate was evaluated by MTT assay. Results PMA preincubation of the cells significantly enhanced the activity of the total PKC and the membrane fraction, but lowered the PKC activity of the cytosol fraction, as compared with the cells without PMA treatment （P〈0.01）. PKCa expression was upregulated in the membrane fraction and down-regulated in the cytosol fraction in KBV200 cells after PMA preincubation. PKCβ expression was slightly elevated in the cytosol fraction but exhibited no obvious changes in the membrane fraction after PMA pretreatment of the cells. The values of IC50 of vincristine and adriamycin in PMA-treated cells were increased to 2275.5 nmol/L and 233.25 nmol/L, respectively （P〈0.01）. Conclusion PMA can increase the multidrug resistance of KBV200 cells, which suggests the possible involvement of PKC in the mechanism ofmultidrug resistance of tumor cells.

关 键 词：蛋白激酶C 多药耐药 长春新碱 阿霉素 佛波酯 

分 类 号：R73-3[医药卫生—肿瘤]