小檗碱增强K562／DOX细胞对多柔比星敏感性的研究

Effect of Berberine on Sensitivity Enhances of K562/DOX Cells to Doxorubicin

目的探讨小檗碱增强耐药K562/DOX细胞对化疗药多柔比星的敏感性的作用。方法四甲基偶氮唑蓝法检测小檗碱的细胞毒性及其对多柔比星抗肿瘤活性的增强作用;高内涵活细胞成像系统检测无毒剂量小檗碱作用后,多柔比星在K562/DOX细胞内的蓄积量;PI/Hoechst33342双染法检测小檗碱对多柔比星诱导的K562/DOX细胞凋亡的影响;罗丹明123蓄积实验检测小檗碱对P-糖蛋白外排功能的影响。结果 1μmol·L-1为小檗碱的无毒剂量,在此无毒剂量下,小檗碱使多柔比星对K562/DOX细胞的IC50降低了1.5倍;1μmol·L-1小檗碱可使多柔比星在K562/DOX细胞内的蓄积量增加,增强多柔比星诱导的K562/DOX细胞凋亡,增加K562/DOX细胞内罗丹明123的蓄积量,从而抑制P-糖蛋白的外排功能。结论小檗碱可通过抑制K562/DOX细胞膜上P-糖蛋白的外排功能,增加K562/DOX细胞内多柔比星浓度,促进多柔比星对耐药细胞的诱导凋亡作用,逆转K562/DOX细胞的多药耐药性。

OBJECTIVE To investigate berberine＇s potential to enhance the sensitivity of doxorubicin-resistant human leukemia cell lines （K562/DOX） to doxorubicin in vitro. METHODS MTT assay was performed to determine the effect of berberine on the sensitivity of K562/DOX cells to doxorubicin. Doxorubiein accumulation assay was performed by Arrary Scan VTr HCS600 High-Contents. The effect of berberine on doxorubicin-induced cell apoptosis was tested by PI/Hoechst 33342 assay. The efflux activity of P-gp was investigated by measuring the accumulation of the rhodamine 123 after treatment with berberine. RESULTS 1 μm01·L^-1 of berberine, as a wild dose, reduced IC50 of doxorubicin to K562/DOX ceils by 1.5 times. 1 μm01·L^-1 of berberine showed the enhancement effect on doxorubicin-induced apoptosis. The increase of doxorubicin and rhodamine 123 accumulation was observed in K562/ DOX cells treated with l μm01·L^-1 of berberine which indicated the inhibition of the activity of P-gp. CONCLUSION Berberine is shown to effectively enhance chemosensitivitv of K562/DOX cells to Dox by inhibiting the efflux activity of P-gp.