羽扇豆醇抑制乳腺癌 MCF-7增殖迁移和侵袭及机制研究

Inhibitory Effects and Mechanisms of Lupeol on Proliferation, Migration and Invasion of Human Breast Cancer MCF-7 Cells

目的 探讨羽扇豆醇抑制乳腺癌MCF-7增殖迁移和侵袭及可能机制。方法 以人乳腺癌MCF-7细胞为对象,根据加入羽扇豆醇的浓度不同分为不同组(羽扇豆醇最终质量浓度分别为0、7.5、15、30、60、90 mg/L),以MTT法检测培养24 h后细胞增殖情况,划痕实验检测细胞迁移情况,Transwell法检测细胞侵袭能力,RT-PCR法检测Wnt/β-catenin信号通路下游基因表达,蛋白免疫印迹法检测细胞周期蛋白D1(cyclin-D1)、基质金属蛋白酶2(MMP-2)、C-myc、β-联蛋白(β-catenin)蛋白表达。结果 不同浓度羽扇豆醇下MCF-7细胞存活率、迁移率及侵袭细胞数量均明显不同,比较差异有统计学意义(P<0.05),与对照组(羽扇豆醇0.00 mg/L,即不加入羽扇豆醇)比较,加入7.5、15、30、60、90 mg/L羽扇豆醇组细胞的存活率、迁移率、侵袭细胞数量均明显下降,差异均有统计学意义(P<0.05);不同浓度羽扇豆醇下MCF-7细胞中Wnt、cyclin-D1、MMP-2、C-myc、β-catenin基因蛋白表达量明显不同,比较差异有统计学意义(P<0.05),与对照组比较,加入7.5、15、30、60、90 mg/L羽扇豆醇组细胞中的Wnt、cyclin-D1、MMP-2、C-myc、β-catenin基因蛋白表达量均明显下降,差异均有统计学意义(P<0.05)。结论 羽扇豆醇对人乳腺癌MCF-7细胞的增殖迁移及侵袭均具有明显的抑制作用,其机制可能是与通过抑制Wnt/β-catenin信号通路的相关基因表达有关。

Objective To investigate inhibitory effects and mechanisms of lupeol on the proliferation, migration and invasion of human breast cancer MCF-7 cells.Methods Taking human breast cancer MCF-7 cells as the research objects, they were divided into different groups according to different concentrations of lupeol(the final mass concentrations of lupeol were 0,7.5,15,30,60,90 mg/L).The proliferation of MCF-7 cells was determined by MTT assay after culturing for 24 hours.The scratch test was used to determine the migration ability of MCF-7 cells.The invasion ability of MCF-7 cells was determined by Transwell.The downstream genes of the Wnt/β-catenin signaling pathway in MCF-7 cells were determined by RT-PCR.The protein expressions of cyclin-D1,matrix metalloproteinase-2(MMP-2),C-myc and β-catenin were determined by Western blot.Results The survival rate, migration rate and invaded cells of MCF-7 cells were significantly different with different doses of lupeol, and the difference was statistically significant(P<0.05).The survival rate, migration rate and invaded cells in 7.5,15,30,60 and 90 mg/L lupeol groups were significantly decreased with compared those of the control group(lupeol 0.00 mg/L,that is without lupanol),and the difference was statistically significant(P<0.05).The protein expressions of Wnt, cyclin-D1,MMP-2,c-myc and β-catenin genes of MCF-7 cells were significantly different with different doses of lupeol, and the difference was statistically significant(P<0.05).The protein expressions of Wnt, cyclin-D1,MMP-2,c-myc and β-catenin genes in 7.5,15,30,60 and 90 mg/L lupeol groups were significantly decreased with compared those of the control group(lupeol 0.00 mg/L,that is without lupanol),and the difference was statistically significant(P<0.05).Conclusion Lupeol can inhibit the proliferation, migration and invasion of human breast cancer MCF-7 cells.The mechanism may be related to the inhibitory of expressions of related genes of Wnt/β-catenin signaling pathway.