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目的探讨吴茱萸碱(evodiamine,EVO)对白血病K562细胞及其耐药株K562/Adr的增殖、细胞周期及多药耐药性(MDR)的影响。方法用细胞增殖毒性检测试剂盒(CCK-8)检测EVO和/或柔红霉素(DNR)对细胞增殖的影响,并计算耐药指数(RI)和逆转倍数(RF);流式细胞仪检测EVO和/或DNR对K562及K562/Adr细胞周期的影响;流式细胞仪检测K562及K562/Adr细胞内DNR的荧光强度;定量PCR检测K562及K562/Adr细胞中MDR1基因的表达;Western blotting检测K562及K562/Adr细胞中MDR1、BCRP蛋白的表达。结果EVO、DNR作用于K562和K562/Adr细胞后,细胞增殖受到抑制,且呈剂量和时间依赖性;与K562细胞相比,K562/Adr细胞对DNR的RI为30.54,K562/Adr细胞对EVO的RI为19.09。当EVO(0.125 gmol/L)与不同浓度DNR联合作用后,能使DNR对K562/Adr细胞的IC_(50)明显下降,DNR+EVO对K562/Adr细胞的RF为12.07;EVO、DNR单独或联合作用于K562及K562/Adr细胞后,能够使K562/Adr细胞BCRP、MDR1蛋白及mRNA表达水平均明显下降。结论EVO能有效逆转白血病K562/Adr细胞对DNR的耐药现象,而这种作用可能与EVO通过减少细胞膜上多药耐药蛋白MDR1的表达有关。
Objective To investigate the effects of evodiamine (EVO) on the proliferation, cell cycle and multidrug resistance (MDR) of K562 / K562 / Leukemia cell line K562 / Adr. Methods The effect of EVO and / or daunorubicin (DNR) on the cell proliferation was detected by CCK-8 kit, and the drug resistance index (RI) and reversal factor (RF) were calculated. Flow cytometry The effect of EVO and / or DNR on the cell cycle of K562 and K562 / Adr cells was examined. The fluorescence intensity of DNR in K562 and K562 / Adr cells was detected by flow cytometry. The expression of MDR1 gene in K562 and K562 / Adr cells was detected by quantitative PCR. blotting was used to detect the expression of MDR1 and BCRP protein in K562 and K562 / Adr cells. Results The proliferation of K562 and K562 / Adr cells was inhibited by EVO and DNR in a dose-and time-dependent manner. Compared with K562 cells, the RI of K562 / Adr cells was 30.54 and that of K562 / Adr cells on EVO The RI is 19.09. The combination of EVO (0.125 gmol / L) and different concentrations of DNR decreased the IC 50 of DNR to K562 / Adr cells, and the RF of DNR + EVO to K562 / Adr cells was 12.07. EVO, DNR alone or When combined with K562 and K562 / Adr cells, the expression of BCRP, MDR1 protein and mRNA in K562 / Adr cells were significantly decreased. Conclusion EVO can effectively reverse the drug resistance of K562 / Adr cells to DNR, which may be related to the decrease of MDR1 expression in EVO cells.