华蟾素对乳腺癌MDA-MB-231细胞增殖迁移凋亡的影响

Effect of Cinobufacini on MDA-MB-231 Cells Proliferation,Migration and Apoptosis

目的:研究华蟾素对MDA-MB-231细胞增殖、迁移和凋亡的影响,并探讨其可能的作用机制。方法:体外培养三阴性乳腺癌细胞MDA-MB-231,用倒置显微镜观察不同浓度华蟾素对MDA-MB-231细胞形态的影响;CCK-8实验检测华蟾素对MDA-MB-231细胞增殖的抑制作用;划痕实验检测华蟾素对MDA-MB-231细胞迁移能力的抑制作用;FITC/PI双染流式细胞术检测不同浓度华蟾素对MDA-MB-231细胞凋亡率的影响;Western Blot检测凋亡相关蛋白Bax、Bcl-2、Cleaved capsase-3、Cleaved caspase-9和PTHrP的表达。结果:CCK-8结果显示华蟾素可以时间、浓度依赖性地抑制MDA-MB-231细胞增殖;划痕实验结果表明华蟾素可抑制MDA-MB-231细胞迁移;流式细胞术结果表明华蟾素可浓度依赖性地诱导MDA-MB-231细胞凋亡;Western Blot结果表明,华蟾素可上调MDA-MB-231细胞中促凋亡蛋白Bax、Cleaved capsase-3、Cleaved caspase-9,下调抑凋亡蛋白Bcl-2。此外,华蟾素还可抑制MDAMB-231细胞中PTHrP的表达。结论:华蟾素通过线粒体途径诱导MDA-MB-231细胞凋亡可能是其抗三阴性乳腺癌的机制;华蟾素通过下调乳腺癌MDA-MB-231细胞中PTHrP进而抑制溶骨性骨转移的发生,有待于后续的动物实验来进行验证。

Objective:To investigate the effect of cinobufacini on MDA-MB-231 cells proliferation,migration and apoptosis,and detect the underlying mechanism.Methods:MDA-MB-231 cells were treated with different concentrations of cinobufacini,the morphological changes of MDA-MB-231 cells were observed under inverted microscope.CCK-8 assay was used to detect the effect of cinobufacini on MDA-MB-231 cell proliferation.Scratches test was employed to investigate cell migration in MDA-MB-231 cells and cell apoptosis was analyzed by flow cytometry with the staining of Annexin V-FITC/PI.Western Blot was employed to detect the expression level of Bax,Bcl-2,Cleaved capsase-3,Cleaved caspase-9 and PTHrP.Results:CCK-8 results showed that cinibufacini could inhibit the proliferation of MDA-MB-231 cells in a concentration and time-dependent manner.Scratches test confirmed that cinibufacini treatment reduced the migration ability of MDA-MB-231 cells.The results of flow cytometry suggested that cinibufacini could induce apoptosis in MDA-MB-231 cells in a concentration dependent manner.The expression levels of Bax,Cleaved capsase-3,Cleaved caspase-9 were increased,whereas the expression levels of Bcl-2 and PTHrP were decreased with the treatment of cinobufacini.Conclusion:Cinobufacini could induce apoptosis through the mitochondrial apoptotic pathway to exert anti-tumor effect on MDA-MB-231 cells.The effect of cinobufacini on breast cancer osteolytic bone metastasis needs to be further studied in vivo.