Objective: To investigate the synergistic action and the potential internal mechanism of Genistein ( Gen ) combined with Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand ( TRAIL ) in proliferation and apoptosis of breast cancer cell line MCF-7.Methods: MCF-7 cells were treated with Gen at doses of 1.25, 2.5, 5, 10, 20 and 40μg/mL and with TRAIL of 1, 10, 100 and 1000 ng/mL, respectively.Cell proliferation was determined by MTT assay.Based on the results of MTT, Gen ( 20 μg/mL ) and TRAIL ( 100ng/mL ) were administered for the follow-up tests.The MCF-7 cells were then divided into four groups: the control group, Gen ( final concentration: 20 μg/mL ) group, TRAIL ( final concentration: 100n g/mL ) group, and the combination group.After different treatment, apoptosis of MCF-7 cells was detected by flow cytometry ( FCM ).Caspase-3 activity was measured by immunofluorescence.And the expression of NF-κB was detected by enzyme-linked immunosorbent assay ( ELISA ).Results: TRAIL-inhibited proliferation and TRAIL-induced apoptosis in MCF-7 cells were promoted by the combined use of Gen.The inhibition ratio reached 63.78％ ± 2.61％ and apoptotic rate was increased to 42.20％ ± 1.35％, significantly higher than thoese in the group treated with TRAIL alome ( P ＜ 0.01 ).In addition, compared with TRAIL-treated cells, cells treated with Gen and TRAIL showed enhanced caspase-3 activity ( 44.000 ± 0.445 μmol/L ).The content of NF-κB ( 177.453 ± 25.389 pg/mL ) was significantly lower in the group treated with the combination therapy than in the group treated with TRAIL alone ( 343.333 ± 8.064 pg/mL ) ( P ＜ 0.01 ).Conclusion: Gen can synergize TRAIL to inhibit cell proliferation and induce apoptosis, thus increasing the sensitivity of TRAIL in MCF-7 cells.The mechanism might be that Gen has a synergistic action with TRAIL to activate caspase-3 and inhibit the formation of NF-κB, resulting in the apoptosis of MCF-7 cells.%目的:研究金雀异黄素是否协同TRAIL诱导乳腺癌MCF-7细胞发生凋亡,并探讨其可能的内在机制.方法:首先,MCF-7细胞分别经过1.25,2.5,5,10,20,40μg/mL金雀异黄素及1,10,100,1000 ng/mL TRAIL单独处理后,应用MTT法检测MCF-7细胞增殖情况,根据其结果选择终浓度为20μg/mL金雀异黄素及100 ng/mL TRAIL作为后续试验的浓度;接着,MCF-7细胞分为4组,即时照组、Gen组(终浓度为20 μg/mL)、TRAIL组(终浓度为100 μg/mL)及Gen+TRAIL组.细胞经不同处理后,流式细胞仪检测细胞凋亡率;免疫荧光法检测细胞凋亡中Caspase-3活性;应用酶联免疫吸附方法检测细胞NF-kB含量.结果:联合应用Gen后,明显增强TRAIL对MCF-7细胞增殖的抑制[抑制率为(63.78±2.61)%],并且促进TRAIL诱导细胞凋亡的发生[凋亡率为(42.20±1.35)%],均分别高于相应的单独TRAIL处理组(P＜0.01).另外,TRAIL单独处理的MCF-7细胞Caspase-3活性为17.324±0.880μmol/L/hr/mg protein,NF-κB的含量为343.333±8.064 pg/mL;而在联合应用Gen以后,Caspase-3活性增高(44.000±0.445μmol/L/hr/mg protein),同时NF-κB的合成受到抑制(177.453±25.389 pg/mL),与相应单独用药组比较差异具有统计学意义(P＜0.01).结论:金雀异黄素可协同TRAIL诱导乳腺癌MCF-7细胞发生凋亡、增加乳腺癌细胞对TRAIL的敏感性,其可能的机制是Gen协同TRAIL激活了细胞凋亡过程中Caspase-3,并进一步抑制了NF-κB的合成,从而最终导致乳腺癌MCF-7细胞凋亡的发生.
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