三羟异黄酮与化疗药物联合作用对人乳腺癌细胞株MDA-MB-453增殖的影响

Effects of genistein and chemotherapeutic agents on proliferation of human breast cancer cell line MDA-MB-453

目的观察三羟异黄酮（genistein,GEN）与紫杉醇（paclitaxel,PTX）或阿霉素（doxorubicin,DOX）联合作用对体外培养的HER2/neu高表达人乳腺癌细胞株MDA-MB -453增殖的作用,探讨三羟异黄酮和化疗药物的联合抗癌效应.方法 GEN和化疗药物PTX、DOX单独或联合处理体外培养的人乳腺癌细胞MDA-MB-453,MTT法测定细胞增殖抑制率并用联合用药公式分析合并效应,流式细胞仪分析细胞周期,形态学观察和Annexin-V-FITC/PI双标记法检测细胞凋亡.结果 GEN、PTX、DOX单独作用于乳腺癌MDA-MB-453细胞,均可抑制细胞增殖,引起细胞周期阻滞,并诱导细胞凋亡.10、20 μmol/L的GEN与DOX联合作用时,q值在0.85～1.15之间,二者的联合效应为相加效应;40 μmol/L的GEN与DOX联合作用时,q＞1.15,二者的联合效应为协同效应;而各剂量的GEN与PTX联合作用时,q＜0.85,二者的联合效应为拮抗作用.GEN（40 μmol/L）可增强DOX诱导MDA-MB-453细胞凋亡的作用,而削弱PTX诱导MDA-MB-453细胞凋亡的作用.结论 GEN能增加或协同DOX对MDA-MB-453细胞的抑制增殖作用,而拮抗PTX的抑制增殖作用.

Objective To investigate the combined effects of genistein and chemotherapeutic agents on the growth of breast cancer cell MDA-MB-453 in vitro. Methods HER-2/neu-overexpressing MDA-MB-453 breast cancer cells were treated by genistein or doxorubicin or paclitaxel alone, or genistein and doxorubicin, or genistein and paclitaxel in vitro. Cell proliferation was measured by MTT assay. Distribution of cell cycle was determined by flow cytometry. Cell apoptosis was observed by light microcopy and flow cytometry with Annexin-V-FITC/PI dual labelling. The combined effect of two drugs was evaluated by q value, less than 0. 85, in the range of 0. 85 and 1.15, more than 1.15, which respectively meant that the combined effect of the drugs is antagonistic, additive and synergistic. Results Genistein, doxorubicin, paclitaxel alone inhibited the proliferation, blocked the cell cycle and induced apoptosis of human breast cancer cell MDA-MB453 line in vitro. The inhibitory rate （IR） of genistein （10, 20μmol/L） combined with doxorubicin was higher than that of the drug v used alone, and q value was between 0. 85 and 1. 15, suggesting the combined effect is additive; IR of genistein （40 μmol/L） combined with doxorubicin was higher than that of the drug used alone, and q value was more than 1.15, suggesting the combined effect is synergetic ; IR of genistein combined with paclitaxel was lower than that of the drug used alone, which meant the combined effect is antagonistic. Genistein （40μmol/L） promoted apoptosis induced by doxorubicin, but attenuated apoptosis induced by paclitaxel in MDA-MB-453 cells. Conclusion The effect of genistein combined with doxorubicin is additive or synergetic on the growth of MDA-MB-453, whereas it is antagonistic when combined with paclitaxel.