白藜芦醇四甲氧基衍生物提高耐药人乳腺癌细胞MCF-7/DOX对阿霉素的敏感性

Resveratrol analog,3,5,2',4'-Tetramethoxystilbene,enhance the efficacy of MCF-7/DOX to doxorubicin

目的:研究白藜芦醇四甲氧基衍生物(TMS)对人乳腺癌细胞MCF-7和耐阿霉素(doxorubicin,DOX)的人乳腺癌细胞MCF-7/DOX这两株细胞增殖的影响,探讨白藜芦醇四甲氧基衍生物能否提高耐药细胞MCF-7/DOX对阿霉素的敏感性。方法:用SRB法检测白藜芦醇四甲氧基衍生物的细胞毒性,同时检测白藜芦醇四甲氧基衍生物和DOX联用后对细胞株MCF-7/DOX药物敏感性的影响,用Western-blot方法检测不同浓度的白藜芦醇四甲氧基衍生物对P-gp蛋白表达的影响,罗丹明转运实验考察白藜芦醇四甲氧基衍生物对P-gp转运活性的影响。结果:白藜芦醇四甲氧基衍生物(7.5、15、30μg/ml)能够有效抑制耐药细胞MCF-7/DOX的增殖,然而与此相应的浓度范围内白藜芦醇的细胞增殖抑制作用较差,使用非细胞毒性剂量(3、6μg/ml)的白藜芦醇四甲氧基衍生物能有效提高MCF-7/DOX对阿霉素的敏感性,Western-blot结果显示6μg/ml的白藜芦醇四甲氧基衍生物能够明显下调P-gp的表达,此外,罗丹明转运实验显示白藜芦醇四甲氧基衍生物组(3、6μg/ml)细胞内罗丹明123积累量均有上升,表明白藜芦醇四甲氧基衍生物可以抑制P-gp的转运活性。结论:白藜芦醇四甲氧基衍生物可通过抑制P-gp表达和活性的方式增加MCF-7/DOX对阿霉素的敏感性,从而逆转耐药。

Objective： To study the effect of 3,5,2,4ZTetramethoxystilbene on parental human breast cancer MCF-7 ceils and resistance MCF-7/ DOX cells, and explore if 3,5,2,4＇-Tetramethoxystilbene can improve the sensitivity of resistant cells to doxorubicin. Methods： The effects of doxorubicin （DOX） and 3,5,2＇,4＇-Tetramethoxystilbene on cell viability were assessed by the SRB assay, the protein expression of P-gp were determined by western-blot, and the P-gp transporter activity were investigated by the rhodamine 123 （ Rh123 ） accumulation assay. Re- suits： Cytotoxicity assays showed that 3,5,2,4＇-Tetramethoxystilbene （7.5, 15 and 30 μg/ml） significantly inhibited cell growth. The inhi- bition effects of 3,5,2,4＇-Tetramethoxystilbene were greater than that of resveratrol. SRB assay showed that 3,5,2,4＇-Tetramethoxystilbene at the concentration of 3 and 6 μg/ml sensitized MCF-7/DOX to doxorubicin compared with doxorubiein treatment alone. Western-blot showed that 3,5,2＇, 4＇-Tetramethoxystilbene at the concentration of 6μg/ml suppressed P-gp expression. Moreover Rh123 transport experiments showed that （ 3 and 6 μg/ml） 3,5,2,4＇-Tetramethoxystilbene increased the intracellular accumulation of Rh123. Conclusion： 3,5,2,4 ZTet- ramethoxystilbene can enhance the sensitivity of MCF-7/DOX to doxorubicin by inhibiting P-gp expression and activity.