二氢杨梅素对K562/A02细胞阿霉素耐药性的逆转作用  

Reversal Effect of Dihydromyricetin on Drug Resistance of K562/A02 Cell Line to Adriamycin

作　　者：李碧蓉[1] 王乐[1] 韩维娜[1] 夏琳钦 唐姝[2] 

机构地区：[1]邵阳医学高等专科学校生理学教研室,湖南邵阳422000 [2]邵阳医学高等专科学校病理学教研室,湖南邵阳422000

出　　处：《中国实验血液学杂志》2016年第5期1369-1374,共6页Journal of Experimental Hematology

基　　金：邵阳市科技计划项目(N1228);湖南省教育厅科研项目(09C047)资助;湖南省教育厅科研项目(13C868)

摘　　要：目的:探讨二氢杨梅素(dihydromyricetin,DMY)对人慢性粒细胞白血病耐药细胞株K562/A02细胞对阿霉素(adriamycia,ADM)耐药的逆转作用及可能机制。方法:用DMY(5、10、20、40、60、80和100 mg/L)和ADM((0.05-100 mg/L)处理K562和K562/A02细胞48 h,采用MTT法检测细胞活力及DMY对K562/A02细胞ADM耐药的逆转效应,流式细胞术检测细胞内阿霉素的相对浓度,Western blot检测耐药相关基因的P-糖蛋白(P-glycoprotein,P-gp)、多药耐药相关蛋白(multidrug resistance related protein 1,MRP1)、谷胱甘肽转移酶π(glutathione transferaseπ,GSTπ)和BCL-2蛋白表达。结果:DM Y能抑制K562和K562/A02细胞的增殖且呈剂量依赖效应(r1=0.37,r2=0.38),IC_(50)分别为71.23±6.51和72.88±5.49 mg/L。5、10和20 mg/L为DMY的低细胞毒性剂量。DMY(5、10和20 mg/L)增敏K562细胞和K562/A02细胞对ADM耐药性且呈剂量依赖效应(r1=-0.62,r2=-0.71),耐药逆转倍数介于1.38-28.59之间。DM Y(5,10和20 mg/L)增加K562/A02细胞内ADM的浓度也呈剂量依赖效应(r=0.34)。与对照组比较,DMY(5,10和20 mg/L)均显著降低了K562/A02细胞中Pgp、M RP1、GSTπ和BCL-2蛋白表达,呈剂量依赖性(r1=-0.41,r2=-0.37,r3=-0.58,r4=-0.46)。与ADM组比较,DM Y(5,10和20 mg/L)+ADM组P-gp、M RP1、GSTπ和BCL-2蛋白表达均显著降低(r1=-0.55,r2=-0.41,r3=-0.38,r4=-0.44)。结论:DM Y可以逆转K562/A02细胞对ADM的耐药性,其作用机制可能与下调耐药相关基因P-gp、MRP1、GSTπ和BCL-2的表达有关。Objective：To investigate the reversal effect of dihydromyricetin（DMY） on drug resistance of K562/A02 cells to adriamycin and explore its possible mechanism.Methods：K562 and K562/A02 cells were treated with DMY（5,10,20,40,60,80 and 100 mg/L） and ADM（100-0.05 mg/L） for48 h.The viability of K562 cells and K562/A02 cells was tested and the reversal effect of DMY on drug resistance of K562/A02 cells to adriamycin was analyzed by MTT.The relative concentration of ADM in cells was measured by flow cytometry.Protein expressions of drug resistance related genes including P-glycoprotein（P-gp）,multidrug resistance associated protein 1（MRP1）,glutathione transferase π（GSTπ） and BCL-2 were measured by Western Blot.Results：The proliferation of K562 and K562/A02 cells was significantly decreased by DMY in dose-dependent manner as compared with control group（r1=0.37,r2=0.38）.The IC50 of ADM on K562 and K562/A02 cells were 71.23 ±6.51 and 72.88 ±5.49 mg/L respectively.DMY（5,10 and 20 mg/L） was low cytotoxicity.DMY（5,10 and 20 mg/L） enhanced the sensitivity of K562/A02 cells to ADM in dose-dependent manner（r1=-0.62,r2=-0.71） and the reversal multiples was from1.38 to 28.591.The relative concentrations of ADM in K562/A02 of DMY（5,10 and 20 mg/L） group cells were significantly increased in dose-dependent manner compared with the control group（r= 0.34）.Compared with the control group,the expressions of drug resistance related protein P-gp,MRP1,GSTπ and BCL-2 were significantly decreased in dose-dependent manner in DMY（5,10 and 20 mg/L） group（r1=-0.41,r2=-0.37,r3=-0.58,r=-0.46）.Compared with the ADM group,the protein expressions of drug resistance related genes P-gp,MRP1,GSTπ and BCL-2 in DMY（5,10 and 20 mg/L） ＋ ADM group were significantly decreased in dose-dependent manner（r1=-0.55,r2=-0.41,r3=-0.38,r4=-0.44）.Conclusion：DMY enhances the sensitivity of K562/A02 cells to ADM,its mechanism may be related with decrease of P-gp,MRP1,GSTπ and BCL-2 expressions

关 键 词：二氢杨梅素 K562/A02细胞 阿霉素耐药 耐药相关基因 

分 类 号：R733.72[医药卫生—肿瘤] R979.1[医药卫生—临床医学]