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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 study the expression of protein kinase C (PKC) in multidrug-resistant KBV200 cells and to explore the relationship between the upregulation of PKC and multidrug resistance in tumor. Methods KBV200 cells were divided into control group and phorbol ester (PMA) group. PMA group was pre-incubated with 200nmol / L PMA, while the control group did not. The 32P incorporation method was used to determine the PKC activity of multidrug-resistant KBV200 cells. The expression and subcellular distribution of PKC subtypes were detected by Western blotting. Cell resistance was detected by MTT assay. Results Preincubation with PMA increased PKC activity and PKC activity in KBV200 cells and decreased PKC activity (P <0.01). Preincubation with PMA increased the expression of PKCα in the membrane fraction, decreased the expression of PKCα in the plasma fraction, and showed no significant change in the membrane fraction PKCβ. The expression of PKCβ in the plasma fraction increased slightly. PMA increased the IC50 of vincristine and doxorubicin in KBV200 cells (P <0.01). Conclusion PMA increases the drug resistance of KBV200 cells, which may be related to the up-regulation of PKC expression.