白藜芦醇苷对乳腺癌细胞转移潜能影响观察

Polydatin inhibits metastasis of human breast cancer and underlying mechanisms

目的探讨白藜芦醇苷对乳腺癌细胞的迁移及侵袭能力的影响并初步探讨其可能的分子机制。方法采用MTT法检测白藜芦醇苷对乳腺癌细胞的生长抑制作用,采用划痕实验及Tranwell细胞迁移实验观察白藜芦醇苷对乳腺癌细胞迁移能力的影响,Tranwell细胞侵袭实验观察白藜芦醇苷对乳腺癌细胞侵袭能力的影响,免疫印迹法检测白藜芦醇苷对乳腺癌细胞内转移相关蛋白表达变化的影响。组间比较采用one-way ANOVA和Dunnett法。结果0~7.0μmol/L白藜芦醇苷处理乳腺癌细胞MDA-MB-231 48h后,细胞存活率随着白藜芦醇苷浓度的增加而降低,差异有统计学意义,F=38.394,P〈0.001;同法处理MDA-MB-468细胞株,结果与MDA-MB-231一致,F=107.748,P〈0.001。细胞迁移实验结果显示,与药物未处理组相比,0.5和1.0μmol/L白藜芦醇苷处理细胞8h后,MDA-MB-231细胞的迁移率显著降低（F=14.461,P=0.005）,分别降低了（11.81±0.06）%（P=0.018）和（16.68±0.04）%（P=0.004）;MDA-MB-468细胞的迁移率也明显下降（F=42.272,P〈0.001）,分别下降了（11.44±0.68）%（P=0.006）和（19.77±0.41）%（P〈0.001）。细胞侵袭实验结果显示,与药物未处理组相比,白藜芦醇苷处理MDA-MB-231细胞12h后,细胞侵袭率明显下降,F=105.793,P〈0.001。0.5μmol/L白藜芦醇苷使MDA-MB-231细胞的侵袭率下降了（43.42±0.54）%,P〈0.001;1.0μmol/L藜芦醇苷使MDA-MB-231细胞的侵袭率下降了（41.92±0.46）%,P〈0.001。白藜芦醇苷处理MDA-MB-468细胞16h后,细胞的侵袭率也明显降低,F=136.234,P〈0.001。0.5和1.0μmol/L白藜芦醇苷使MDA-MB-468细胞的侵袭率分别降低了（36.78±0.39）%（P〈0.001）和（38.89±0.86）%（P〈0.001）。蛋白质印迹实验提示,白藜芦醇苷对乳腺癌细胞迁移及侵袭能力的抑制作用可能与白藜芦醇苷抑制N-Cadherin及增加E-Cadherin的表达有关。结论白藜芦醇苷可在体外抑制乳腺癌的迁移及侵袭能力,具有应用于乳腺癌临床治疗的潜能。

OBJECTIVE To investigate the effect of Polydatin on the ability of metastasis in human breast cancer cell lines. METHODS Cell proliferation was measured by MTT viability assay after being treated with polydatin at dif- ferent concentrations. Wound healing assay and Transwell cell migration assay were employed to investigate the ability of cell migration. Cell invasion ability was evaluated by Transwell cell invasion assay. The expression level of E-Cadherin, N-Cadherin,β-catenin,a-catenin, MMP-2 and MMP-9 protein, which was associated with apoptosis, were measured by Western blot analysis. RESULTS After breast cancer MDA-MB-231 ceils were incubated with 0-7.0 μmol/L poldatin for 48 h, cell viability decreased in a dose dependent manner. The difference was statistically significant （F= 38. 394, P〈0.001）. Similar results were seen in MDA-MB-468 ceils when ceils were treated with polydatin（F= 107. 748, P〈0. 001）. Polydatin inhibited proliferation of breast cancer ceils in a dose-dependent manner. In comparison with thecontrol, reduction of cell migration was seen when cells were incubated with polydatin for 8 h. The inhibition percentage of 0.5μmol/L and 1.0 μmol/L polydatin was （11.81±0.06）%（P=0. 018） and （16.68±0.04）% （P=0. 004）in breast cancer MDA-MB-231 cells（F=14. 461,P=0. 005） and （11.44±0.68）%（P=0. 006） and （19.77±0.41）%（P〈0. 001） in breast cancer MDA-MB-468 cells（F=42. 272,P〈0. 001）. In the invasion assay, reduction percentage of 0.5 μmol/L and 1.0 μmol/L polydatin on cell invasiveness was （43.42±0.54）% （P〈0. 001） and （41.92± 0.46）% （P〈0. 001）in MDA-MB-231 cells after cells were treated for 12 h（F= 105. 793,P〈0. 001） and （36.78±0. 39）% （P〈0. 001） and （38.89±0.86）%（P〈0.001）inMDA-MB-468 cells after cells were treated for 16 h（F=136.234,P〈0.001）. Western blot analysis indicated that polydatin down-regulated expression of N-Cadherin and up-regulate expression of E Cadherin. Simultaneously expression of MMP-2 and MMP-9 protein was not affected. CONCLUSIONS polydatin is a promising anti-metastasis agent in breast cancer in vitro. It has the potential to be useful in breast cancer treatment.