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目的探讨蛋白激酶C(PKC)活性、亚细胞分布与KBV200细胞株多药耐药(MDR)的关系。方法MTT法检测KB和KBV200细胞对细胞毒药物的耐药性,32P掺入法检测两株细胞的PKC活性。结果长春新碱(VCR)、阿霉素(ADR)对KBV200细胞的半数抑制浓度(IC50)值分别是KB细胞的65.03和19.8倍;KBV200细胞的膜组分、胞质组分的PKC活性均高于KB细胞(P<0.01),KBV200细胞PKC总活性是KB细胞的2.12倍;佛波酯可升高KBV200细胞总或膜组分PKC活性,VCR、ADR的IC50值升高(P<0.01);十字孢碱可降低KBV200细胞两种组分PKC活性,VCR、ADR的IC50值降低(P<0.01),对VCR、ADR的逆转倍数分别是5.46和1.96倍。结论PKC与KBV200细胞株的MDR表型关系密切,PKC可能介导了KBV200细胞的MDR。
Objective To investigate the relationship between protein kinase C (PKC) activity, subcellular distribution and multidrug resistance (MDR) in KBV200 cells. Methods MTT assay was used to detect the drug resistance of KB and KBV200 cells. The 32P incorporation method was used to detect the PKC activity of the two cells. Results The median inhibitory concentration (IC50) of vincristine (VCR) and doxorubicin (ADR) on KBV200 cells were 65.03 and 19.8 times higher than that of KB cells respectively. The membrane fraction and cytoplasmic fraction of KBV200 cells were both PKC (P <0.01). The total PKC activity of KBV200 cells was 2.12 times that of KB cells. Phorbol ester increased the total or membrane-derived PKC activity of KBV200 cells. The IC50 values of VCR and ADR increased (P <0.01) ). Staurosporine decreased the PKC activity of both components in KBV200 cells. The IC50 values of VCR and ADR decreased (P <0.01). The reversal multiples of VCR and ADR were 5.46 and 1.96 times respectively. Conclusion PKC is closely related to the MDR phenotype of KBV200 cells. PKC may mediate the MDR of KBV200 cells.