茯苓酸对小鼠黑色素瘤细胞增殖、迁移和侵袭的影响及机制研究

The Effect of Pachymic Acid on the Proliferation, Migration and Invasion of Mouse Melanoma Cells and Study on the Mechanism

目的:研究茯苓酸(pachymic acid,PA)对小鼠黑色素瘤B16细胞增殖、迁移和侵袭的影响及其可能机制。方法:采用CCK8法和细胞克隆形成实验检测不同浓度(20、40、80μmol/L)PA对B16细胞增殖的影响;细胞划痕和Transwell小室实验观察PA对B16细胞迁移和侵袭能力的影响;Western Blot法和免疫荧光法检测PA对B16细胞的上皮-间质转化相关蛋白表达的变化。结果:PA可以明显抑制B16细胞增殖,并呈一定的浓度依赖性关系(P<0.01);PA干预24 h后可以显著抑制B16细胞的迁移和侵袭能力(P<0.01);40μmol/L的PA能上调B16细胞的E-cadherin蛋白表达(P<0.05)并下调Vimentin蛋白的表达(P<0.05);免疫荧光法检测显示40μmol/L的PA可明显降低B16细胞Vimentin蛋白的表达。结论:PA可以抑制B16细胞的增殖、迁移和侵袭,其机制可能与上调B16细胞的E-cadherin蛋白表达,降低Vimentin蛋白表达,从而抑制B16细胞的上皮-间质转化有关。

Objective:To study the effects of pachymic acid(PA)on the proliferation,migration and invasion of mouse melanoma B16 cells and its possible mechanisms.Methods:CCK8 assay and cell clone formation assay were used to detect the effects of different concentrations(20,40,80μmol/L)of PA on the proliferation of B16 cells;cell scratch and Transwell assays were used to observe the effects of PA on the migra-tion and invasive ability of B16 cells;and Western Blot and immunofluorescence assays to detect the effects of PA on the expression of epithelial-mesenchymal transition-related protein of B16 cells.Results:PA could obviously inhibit the proliferation of B16 cells in a concentration-dependent manner(P<0.01);PA could remarkably inhibit the migration and invasion ability of B16 cells after 24 h of intervention(P<0.01);40μmol/L PA up-regulated the expressions of E-cadherin protein(P<0.05)and down-regulated the expression of Vimentin protein of B16 cells(P<0.05);immunofluorescence assay showed that 40μmol/L PA apparently decreased the expression of Vimentin protein in B16 cells.Conclusion:PA can inhibit the proliferation,migration and invasion of B16 cells,and its mechanism may be related to the up-regulation of E-cadherin protein expression and the reduction of Vimentin protein expression in B16 cells,thus inhibiting the epithelial-mesenchymal transition of B16 cells.