诃子乙醇提取物调控SOX9对乳腺癌细胞增殖、凋亡、周期、迁移和侵袭的分子机制研究

Study on the mechanism of terminalia chebula extract regulating SOX9 to inhibit the proliferation,apoptosis, cycle, migration and invasion of breast cancer cells

目的探讨诃子乙醇提取物(TCE)通过调控高迁移率族盒蛋白9(SOX9)对乳腺癌细胞增殖、凋亡、周期、迁移和侵袭的分子机制研究。方法通过MTT法检测细胞活力;流式细胞术实验检测细胞凋亡和细胞周期;通过Western blot检测p21、Bcl-2、Bax和SOX9蛋白表达水平;Transwell小室检测细胞的迁移和侵袭能力;通过qRT-PCR检测SOX9 mRNA相对表达量。结果随着TCE浓度的增加,细胞活力、Bcl-2蛋白表达、S期细胞比例、迁移细胞数目和侵袭细胞数目逐渐降低,细胞凋亡率、p21、Bax蛋白表达和G0/G1期细胞比例逐渐增加。TCE组的SOX9 mRNA相对表达量和蛋白表达均低于Con组。si-SOX9组的SOX9 mRNA相对表达量及蛋白表达、细胞活力、Bcl-2蛋白表达、S期细胞比例、迁移细胞数目和侵袭细胞数目低于si-NC组;而细胞凋亡率、p21、Bax蛋白表达和G0/G1期细胞比例均高于si-NC组。TCE+SOX9组的SOX9 mRNA相对表达量及蛋白表达、细胞活力和Bcl-2蛋白表达、S期细胞比例、迁移细胞数目和侵袭细胞数目均高于TCE+pcDNA组,而细胞凋亡率、p21和Bax蛋白表达及G0/G1期细胞比例均低于TCE+pcDNA组。结论 TCE可以调控SOX9表达抑制MCF-7细胞的增殖、迁移和侵袭,并促进其凋亡,使细胞周期阻滞。

Objective To investigate the molecular mechanism of terminalia chebula extract(TCE) inhibiting the proliferation, apoptosis cycle, migration and invasion of breast cancer cells by regulating SRY-related high-mobility group box 9(SOX9). Methods Cell activity was detected by MTT assay;cell apoptosis and cell cycle were measured by flow cytometry;the protein expression level of p21, Bcl-2, Bax and SOX9 were detected by Western blot;Transwell assay detected the ability of cell migration and invasion;the relative expression of SOX9 mRNA was detected by qRT-PCR. Results With the increase of TCE concentration, cell viability, Bcl-2 protein expression, S-phase cell ratio, number of migrating cells, and number of invasive cells gradually decrease, and cell apoptosis rate, p21, Bax protein expression, and G0/G1 phase cell ratio gradually increase. The relative expression of SOX9 mRNA and protein expression in TCE group were lower than those in Con group.The relative expression of SOX9 mRNA and protein expression, cell viability, Bcl-2 protein expression level, S phase cell ratio,number of migrating cells and number of invasive cells in the si-SOX9 group were lower than those in the si-NC group;while the apoptosis rate, p21, Bax protein expression and G0/G1 phase cell ratio were higher than si-NC group. The relative expression of SOX9 mRNA and protein expression, cell viability and Bcl-2 protein expression, S phase cell ratio, number of migrating cells and number of invasive cells in the TCE+SOX9 group were higher than those in the TCE+pcDNA group, while the apoptosis rate, p21 and Bax protein expression and the ratio of G0/G1 phase cells were lower than TCE+pcDNA group. Conclusion TCE can regulate the expression of SOX9, inhibit the proliferation, migration and invasion of MCF-7 cells, promote their apoptosis and block the cell cycle.