吴茱萸碱逆转K562/Adr细胞多药耐药的实验研究  

Experimental study of evodiamine on reversing multidrug resistance of K562/Adr cells

作　　者：李晓朋[1] 冯子强[1] 石雪萍[1] 李静 

机构地区：[1]重庆医科大学基础医学院,组织学与胚胎学教研室,干细胞与组织工程研究室,重庆400016

出　　处：《中草药》2016年第12期2123-2129,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(31271368)

摘　　要：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 study the effect of evodiamine （EVO） on the proliferation, cell cycle, and multidrug resistance （MDR） of leukemia K562 and K562/Adr cells. Methods The effect on the proliferation was detected by CCK-8 kit and/or daunorubicin （DNR）. The resistance index （RI） and reversal fold （RF） were calculated. The effect of EVO on cell cycle was tested by flow cytometry; Flow cytometry was used to check the fluorescence intensity of DNR. The expression ofMDR1 gene in leukemia K562 and K562/Adr cells was detected by q-PCR. The expression of MDR1 and BCRP proteins in leukemia K562 and KS62/Adr cells were detected by Western blotting. Results CCK-8 test results showed that the proliferation ofK562 and K562/Adr cells induced by EVO and DNR was inhibited in a dose and time dependent manner. The RI of DNR on K562/Adr cells was 30.54, the RI of EVO on K562/Adr cells was 19.09, compared with K562 cells; The RF of DNR ＋ EVO on K562/Adr cells was 12,07. The expression of BCRP and MDR1 protein of K562/Adr cells was significantly decreased in K562/Adr cells induced by EVO and DNR. Conclusion Therefore EVO can effectively reverse the DNR resistance in K562/Adr cells, which may be related to reduce the expression of MDR1 on cell membrane.

关 键 词：吴茱萸碱 K562/Adr细胞 多药耐药性 MDR1 BCRP 

分 类 号：R285.5[医药卫生—中药学]