三羟异黄酮对人乳腺癌MCF-7/ADM细胞体外抑瘤效应、细胞周期及凋亡的影响

Genistein Plays Antitumor Role through Cell Cycle and Apotosis Pathways in Human Breast Cancer Cell Line

目的探讨三羟异黄酮(Genistein GEN)对体外培养人乳腺癌耐药细胞MCF-7/ADM抑瘤作用、细胞周期及细胞凋亡的影响。方法采用MTT法检测GEN单独及联合阿霉素对体外培养人乳腺癌MCF-7/ADM细胞的抑制作用;荧光分光光度法检测GEN对阿霉素在MCF-7/ADM细胞的蓄积作用;用流式细胞仪(FCM)检测细胞周期及凋亡率的变化。结果 GEN单独及联合阿霉素对体外培养MCF-7/ADM细胞均有明显生长抑制作用,GEN单独作用48 h后表现为随时间的延长抑制作用明显增强,当GEN浓度达到60μg/ml时,其抑制作用急剧上升(P<0.01)。联合阿霉素后与对照组相比抑制率明显上升(P<0.01),并随GEN浓度的加大其抑制作用增强,细胞内阿霉素的浓度也随之升高。与对照组相比,细胞周期均有G2/M期阻滞作用,细胞凋亡百分比以联合组最高(P<0.01),G1期前出现典型的亚二倍体凋亡峰。结论 GEN单独及联合阿霉素对体外培养人乳腺癌MCF-7/ADM细胞具有抑瘤增效作用,可以提高阿霉素在MCF-7/ADM细胞内的蓄积、对细胞周期具有G2/M期阻滞作用,显著诱导MCF-7/ADM细胞凋亡,可能是其发挥逆转多药耐药分子生物学机制之一。

Objective To study the antitumor effect of genistein（Genistein GEN） in cultured drug-resistant breast cancer cell line of MCF-7/ADM in vitro,and influences of genistein to cell cycle and apoptosis.Methods Inhibitory effect of GEN alone or combined with doxorubicin on the cultured MCF-7/ADM was detected by MTT assay;the accumulative effect of GEN on doxorubicin in MCF-7/ADM was detected by fluorescence spectrophotometry;and cell cycle and apoptosis rate was detecced by flow cytometry（FCM）.Results Significant inhibitory effect on cultured MCF-7/ADM in vitro was not observed under GEN alone or combined with Doxorubicin 48 h later GEN treated alone,the inhibition increased gradually in time-dependent model.When the concentration of GEN reached 60 μg/ml,inhibition effect was markedly increased（P0.01）.When Doxorubicin was added,the inhibition rate was significant increased compared with the control group（P0.01）,and the inhibition strengthened with the increasing concentration of GEN,concentration of intracellular doxorubicin was also increased.Compared with the control group,the cell cycle were both blocked at G2/M phase,apoptosis was found to be the highest percentage in the combination group（P0.01）,typical hypodiploid apoptotic peak was detected before the G1 phase.Conclusion GEN alone and combined with Doxorubicin had an inhibitory and additive effect on cultured human breast cancer cell line MCF-7/ADM in vitro,it could increase the intracellular accumulation of Doxorubicin and arrest cell cycle at phase G2/M,as well as in inducing significant apoptosis of MCF-7/ADM cells,which may be one of its molecular mechanisms of the reversal of multidrug resistance.