目的 探讨柴胡皂苷-d对人乳腺癌细胞MDA-MB-231增殖和转移的作用及机制.方法 经不同浓度柴胡皂苷-d处理MDA-MB-231细胞后, CCK-8法检测细胞活性, 平板克隆实验检测细胞增殖能力;设定最低柴胡皂苷-d作用浓度的MDAMB-231细胞为实验组, 不加任何处理的MDA-MB-231细胞为空白对照组, 通过3D细胞培养实验、细胞划痕实验、transwell实验检测柴胡皂苷-d对细胞迁移和侵袭的影响;流式细胞实验检测其对细胞周期和凋亡的影响;最后通过实时荧光定量PCR、Western印迹法探讨其对细胞增殖和转移的作用机制.结果 柴胡皂苷-d能呈时间及浓度依赖性抑制MDA-MB-231的细胞活性和增殖能力;柴胡皂苷-d 40μmol/L处理后MDA-MB-231细胞形态发生了明显的上皮性改变, 实验组MDA-MB-231细胞的伤口愈合能力、迁移能力和侵袭能力受到明显抑制 (P＜0.01);流式细胞实验发现, 与对照组相比, 柴胡皂苷-d 40μmol/L处理后MDA-MB-231细胞阻滞于G1期, 细胞凋亡率明显升高;实验组细胞Bcl-2、波形蛋白 (vimentin) 和N-钙黏蛋白 (cadherin) 的mRNA和蛋白表达下调 (P＜0.01), 而P21、P27和E-钙黏蛋白mRNA和蛋白质表达上调 (P＜0.01) .结论 柴胡皂苷-d有可能作为潜在的抗肿瘤药物用于抑制乳腺癌细胞增殖和转移.%Objective To investigate the effect and related mechanism of saikosaponin-d (SS-d) on the proliferation and migration of breast cancer MDA-MB-231 cells. Methods The MDA-MB-231 cells were treated with varying concentrations of SS-d.CCK-8 and colony formation analysis were used to evaluate the proliferation ability of MDA-MB-231 cells;3D cell culture, wound healing assay and Transwell migration and invasion assays were used to analyze the ability of cell migration and invasion;flow cytometry was utilized to analyze the cell cycle and apoptosis;semi-quantitative RT (qRT)-PCR and Western blot were used to explore the action mechanism of SS-d on the cell migration and invation. Results SS-d inhibited the proliferation and colony formation ability of MDA-MB-231 cells, both dose-and time-dependently;migration and invasion abilities of the MB-231 cells were also significantly inhibited by the treatment with SS-d (P＜0.01);the treatment with SS-d caused the significant cell cycle arrest at G1 phase and also significant apoptosis-induction in MDA-MB-231 cells;SS-d could up-regulate the expression of P21, P27 E-cadherin (P＜0.01), but down-regulate the expression of Bcl-2, vimentin and N-cadherin (P＜0.01), all detected in both the m RNA and protein levels. Conclusion SS-d was likely to be a potential antitumor drug for the breast cancer therapy to inhibit the proliferation and metastasis of the cancer cells.
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