Genistein抑制Livin基因对恶性黑色素瘤LiBr细胞凋亡、周期及增殖的影响

Effect of livin gene suppression by genistein on apoptosis, cell cycle and proliferation of malignant melanoma LiBr cells

目的观察genistein抑制Livin基因对恶性黑色素瘤LiBr细胞凋亡、周期及增殖的影响。方法应用RT-PCR检测不同浓度genistein作用LiBr细胞48 h后Livin的表达变化;取最适浓度的genistein作用LiBr细胞后应用Annexin V-FITC/PI双染流式细胞术检测其对细胞凋亡的影响,应用PI单染流式细胞术分析对细胞周期的改变,应用Western blot检测其对凋亡效应蛋白Ccaspase-3表达的影响及应用MTT比色法检测其对细胞增殖的作用。结果 Genistein可增加LiBr细胞早、晚期凋亡率[分别为(27.87±5.38)%和(11.87±3.86)%],诱导LiBr细胞凋亡(P<0.01);引起细胞G0/G1期阻滞[G0/G1=(72.11±5.89)%、S=(14.53±3.47)%、G2/M=(12.36±2.64)%];下调caspase-3蛋白表达及抑制细胞增殖(P<0.01)。结论 Genistein可诱导LiBr细胞凋亡、使细胞增殖周期进展受阻、抑制细胞增殖。

Objective To observe the effect of livin gene suppression by genistein on apoptosis, cell cycle and proliferation of malignant melanoma LiBr ceils. Methods RT-PCR was used to detect the expression of livin mRNA in LiBr cells 48 h after treatment with 40 μmol/L gemstein. Flow cytometry with annexin V-FITC/PI double staining was employed to detect cell apoptosis, and the caspase-3 protein expression in the cells following genistein treatment was assayed using Western blotting. The changes in the cell cycle and proliferation of the cells after genistein treatment were examined with flow cytometry with PI staining and MTT colorimetric assay, respectively. Results Genistein can suppress the expression of Livin Gene （87.94% with 40 μmol/L genistein） and induce the apoptosis of LiBr effectively, both in the early and late phases （27.87±5.38% and 11.87± 3.86% respectively）. LiBr ceils in phase G0/G1 increases notably [G0/G1=（72.11±5.89）% 、S=（14.53±3.47）% 、 G2/M=（12.36_±2.64% ）]. Genistein significantly reduced caspase-3 protein expression and inhibit cell proliferation. Conclusion Genistein can suppress Livin Gene expression, induct LiBr cell apoptosis, hinder cell generation cycle, restrain cell proliferation.