蛇葡萄素对人前列腺癌DU145细胞增殖、迁移和侵袭的影响

Effects of Ampelopsin on Proliferation, Migration and Invasion of Human Prostate Cancer DU145 Cells

目的:探讨蛇葡萄素(AMP)对人前列腺癌DU145细胞增殖、迁移和侵袭的影响,研究其对转化生长因子β(TGF-β)诱导的上皮间质转化(EMT)模型的作用。方法:用不同浓度的AMP处理DU145细胞,采用CCK-8法和克隆形成实验检测细胞增殖情况;Transwell法检测细胞的迁移和侵袭能力;细胞免疫荧光实验表征不同浓度TGF-β处理后细胞内E-cadherin和Vimentin表达情况,确定EMT模型最佳诱导浓度;Western blot法进一步检测加药后EMT模型E-cadherin和Vimentin表达情况。结果:AMP对DU145细胞增殖具有显著的抑制作用,24、48、72 h的IC50分别为35.62、32.71、28.43μg·mL^-1;Transwell实验显示AMP处理后穿膜细胞数明显减少,显著抑制细胞的迁移和侵袭能力;细胞免疫荧光实验表征发现TGF-β诱导后细胞内E-cadherin表达下调,Vimentin表达上调,确定10ng·mL^-1为TGF-β诱导EMT模型的最佳浓度。Western blot法检测显示,与对照组相比,TGF-β组和TGF-β+AMP组的细胞内E-cadherin表达下调,Vimentin表达上调,此外,与TGF-β组相比,TGF-β+AMP组细胞内E-cadherin表达变化无统计学差异,但Vimentin表达降低(P<0.05),表明AMP具有逆转EMT过程的作用。结论:AMP不仅能够抑制人前列腺癌DU145细胞的增殖,还可抑制其迁移和侵袭能力,这可能与逆转EMT过程有关。

Objective: To investigate the effects of ampelopsin(AMP) on proliferation, migration and invasion of human prostate cancer DU145 cells, and study the roles of AMP on growth factor-β(TGF-β)-induced epithelial-mesenchymal transition(EMT). Methods: DU145 cells were treated with varying concentrations of AMP, and CCK-8 assay and clone formation assay were utilized to assess cell proliferation. Cell migration and cell invasion abilities were examined by transwell assays. E-cadherin and Vimentin expression of DU145 cells after treatment with different concentrations of TGF-β were detected with immunofluorescence assay to determine the best induction concentration of TGF-β. Western blot analysis was used to measure the protein expression of E-cadherin and Vimentin. Results: AMP significantly inhibited the proliferation of DU145 cells, and its IC50 after 24, 48 and 72 h were 35.62, 32.71 and 28.43 μg·mL^-1, respectively. Transwell invasion assay showed that the numbers of cells across the membrane in the groups adding AMP decreased significantly, which revealed that AMP significantly suppressed the invasion and metastasis ability of the DU145 cells. Immunofluorescence demonstrated that TGF-β increased the expression of E-cadherin and decreased that of Vimentin, at the optimal concentration of 10 ng·mL^-1. The results from the Western blot analysis demonstrated that compared with the control group, the expression of E-cadherin in TGF-β group and TGF-β+AMP group were down regulated, while Vimentin were up regulated.Furthermore, compared with the TGF-β group, the expression of E-cadherin in the TGF-β+AMP group had no significant difference, but the expression of Vimentin decreased(P<0.05), which suggest that AMP may reverse the process of EMT. Conclusions: AMP could suppress the proliferation of human prostate cancer DU145 cells, as well as their migration and invasion abilities, which may be related to the reversal of EMT process.