[关键词]
[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 μmol/L)与不同浓度DNR联合作用后,能使DNR对K562/Adr细胞的IC50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。
[Key word]
[Abstract]
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 of MDR1 gene in leukemia K562 and K562/Adr cells was detected by q-PCR. The expression of MDR1 and BCRP proteins in leukemia K562 and K562/Adr cells were detected by Western blotting. Results CCK-8 test results showed that the proliferation of K562 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.
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[基金项目]
国家自然科学基金资助项目(31271368)