Li Bi-Rong, Wang Le, Han Wei-Na, Xia Lin-Qin, Tang Shu
Department of Physiology, Shaoyang Medical College, Shaoyang 422000, Hunan Province, China.
Department of Pathology, Shaoyang Medical College, Shaoyang 422000, Hunan Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2016 Oct;24(5):1369-1374. doi: 10.7534/j.issn.1009-2137.2016.05.016.
To investigate the reversal effect of dihydromyricetin(DMY) on drug resistance of K562/A02 cells to adriamycin and explore its possible mechanism.
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) for 48 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.
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 IC 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 from 1.38 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).
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.
研究二氢杨梅素（DMY）对K562/A02细胞阿霉素耐药性的逆转作用，并探讨其可能机制。
用DMY（5、10、20、40、60、80和100mg/L）和阿霉素（100 - 0.05mg/L）处理K562和K562/A02细胞48小时。检测K562细胞和K562/A02细胞的活力，并用MTT法分析DMY对K562/A02细胞阿霉素耐药性的逆转作用。通过流式细胞术测量细胞内阿霉素的相对浓度。用蛋白质免疫印迹法检测耐药相关基因包括P-糖蛋白（P-gp）、多药耐药相关蛋白1（MRP1）、谷胱甘肽转移酶π（GSTπ）和BCL-2的蛋白表达。
与对照组相比，DMY以剂量依赖方式显著降低K562和K562/A02细胞的增殖（r1 = 0.37，r2 = 0.38）。阿霉素对K562和K562/A02细胞的半数抑制浓度（IC）分别为71.23±6.51mg/L和72.88±5.49mg/L。DMY（5、10和20mg/L）细胞毒性较低。DMY（5、10和20mg/L）以剂量依赖方式增强K562/A02细胞对阿霉素的敏感性（r1 = -0.62，r2 = -0.71），逆转倍数为1.38至28.591。与对照组相比，DMY（5、10和20mg/L）组K562/A02细胞内阿霉素的相对浓度以剂量依赖方式显著增加（r = 0.34）。与对照组相比，DMY（5、10和20mg/L）组耐药相关蛋白P-gp、MRP1、GSTπ和BCL-2的表达以剂量依赖方式显著降低（r1 = -0.41，r2 = -0.37，r3 = -0.58，r = -0.46）。与阿霉素组相比，DMY（5、10和20mg/L）+阿霉素组耐药相关基因P-gp、MRP1、GSTπ和BCL-2的蛋白表达以剂量依赖方式显著降低（r1 = -0.55，r2 = -0.41，r3 = -0.38，r4 = -0.44）。
DMY增强K562/A02细胞对阿霉素的敏感性，其机制可能与降低P-gp、MRP1、GSTπ和BCL-2的表达有关。
