蛋白激酶C活性、亚细胞分布与KBV200细胞多药耐药性的相关研究

Correlation of protein kinase C activity and its subcellular distrubution with multidrug resistance in KBV200cells

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 investigatetherelationshipbetweenproteinkinaseC(PKC)and multidrugresistance(MDR)in cancercelllineKBV200.Methods MTT assaywas usedto evaluatetheIC 50 of vincristine(VCR)andadriamycin(ADR)in KB celllineanditsVCR-resistantderivativeKBV200cells.PKCactivitiesinthe2celllineswereassayedby measuringthe incorporationof 32 P from[γ- 32 P]ATPintothepeptidesubstrates.Results TheIC 50 valuesof VCRandADRinKBV200cells was64.03and18.8foldsgreaterthanthoseinKB cells.PKCactivitiesof themembraneandcytosolfractioninKBV200cells wereincreasedcomparedwiththatinKB cells,withthetotalPKCactivity1.12-foldhigher.Phorbol-12-myristate-13-acetate increasedPKCactivityof themembranefractionandIC 50 valuesof KBV200cells,whilestaurosporineworkedto theopposite effects.Conclusion PKCmaycontributeto MDRmechanisminKBV200cells.