The effect of elemene on reversal multidurg resistance and the expression of P-glyco-protein in A549/DDP

目的:探讨榄香烯乳(ElEMEnE,ElE)逆转人肺腺癌A549/ddP细胞株耐药性及其机制。方法:采用MTT法检测榄香烯乳单用的细胞毒作用及与顺铂(CISPlATIn,ddP)合用时耐药逆转作用;采用流式细胞术检测榄香烯乳对A549/ddP细胞内罗丹明-123(rHOdAMInE-123,rH123)蓄积的影响;采用WESTErn blOT检测榄香烯乳对耐药细胞A549/ddP细胞膜上P-gP蛋白表达的影响。结果:不同浓度榄香烯对A549/ddP细胞均有一定的抑制作用,呈时间-剂量依赖性效应。单用ddP的IC50为15.46μg/Ml,合用20μg/Ml榄香烯24H,IC50为4.15μg/Ml,逆转耐药倍数为3.63。同时,榄香烯增加A549/ddP细胞内rH123的蓄集,降低细胞膜上PgP的表达,且呈剂量依赖性,表明榄香烯能减少A5 4 9/ddP细胞对药物的外排和抑制P-gP蛋白的表达。结论:榄香烯在体外逆转肿瘤细胞耐药性可能与其抑制P-gP的功能和表达有关。Objective: To investigate the elemene( ELE) reversal of human lung adenocarcinoma A549/DDP cell resistance and its mechanism.Methods: Cytotoxicity of ELE alone or in combination with DDP in A549 / DDP cells was determined by MTT assay.Based on flow cytometric technology,the effect of ELE on the uptake of rhodamine-1 2 3( Rho1 2 3) in A5 4 9 / DDP cells was detected by measuring Rho123 mean fluorescence intensity( MFI).P- gp protein expression on the A549 / DDP cell membrane were detected by Western blot.Results: Different concentration elemene inhibited the A549 / DDP cell proliferation in a time- dose- dependent manner( P < 0.05).Alonely used DDP,IC 50 was 15.46μg / ml; combined 20μg / ml elemene for 24h,DDP IC 50 was 4.15μg / ml,the reversal activity was 3.63 times higher than that of DDP.Elemene increased intracellular accumulation of Rh123 of the A549 / DDP cells, reduced the expression of P- gp on the cell membrane,in a dose- dependent manner,indicating that elemene reduce A549 / DDP cells for drug efflux and inhibition of P- gp protein pump function.Conclusion: The reversal activity of DDP is possible related to inhibition of P- gp function and expression in vitro,which lead to an increased intracellular accumulation of anticancer drugs.福建省自然科学基金面上项目(编号:2010D014