熊果酸对舌癌细胞Tca8113生长、迁移和凋亡的影响及机制研究

Effects of ursolic acid on growth,migration and apoptosis of tongue cancer Tca8113 cells and its mechanism

目的探究熊果酸对人舌癌细胞Tca8113体外生长、迁移和凋亡的影响及机制。方法采用噻唑蓝(MTT)法及克隆形成实验检测熊果酸对Tca8113细胞活性的影响;采用划痕实验检测熊果酸在0、12及24h对Tca8113细胞迁移能力的影响;通过Heochst33342染色和流式细胞术观察熊果酸对Tca8113细胞凋亡的影响;Western blotting检测熊果酸对细胞中磷酸化局部黏着斑激酶(p-Fak)、局部黏着斑激酶(Fak)、活化冬氨酸蛋白水解酶-3(Cleaved caspase-3)、Bcl-2-Associated X的蛋白质(Bax)、B细胞淋巴瘤-2(Bcl-2)蛋白表达的影响。结果熊果酸能抑制Tca8113细胞的活性并促进其凋亡(P<0.05),并呈时间、浓度依赖性。熊果酸抑制Bcl-2,并促进Bax、Cleaved caspase-3表达,促进Tca8113细胞凋亡;能降低p-Fak、Fak的表达,抑制Tca8113细胞的迁移。结论熊果酸能抑制Tca8113细胞的生长及迁移,其机制可能与调节Bcl-2/Bax凋亡通路和抑制Fak的激活相关。

Objective To investigate the effects and mechanism of ursolic acid on the growth and migration of human tongue cancer Tca8113 cells in vitro.Methods The effects of ursolic acid on the viability of Tca8113 were detected by MTT assay and colony formation assay.The effects of ursolic acid on the migration of Tca8113 cells at 0 h,12 h and 24 h were detected by scratch test.Heochst33342 staining and flow cytometry were performed to observe the effects of ursolic acid on the apoptosis of Tca8113 cells.Western blotting was used to detect the effects of ursolic acid on the expression of Bcl-2,Bax,Cleaved Caspase-3,p-Fak and Fak.Results Ursolic acid inhibited the viability of Tca8113 cells and promoted apoptosis in a time-and concentration-dependent manner.The mechanism lies in that ursolic acid increased apoptosis of Tca8113 cells by inhibiting the expression of Bcl-2 and increasing the expression of of Bax,resulting in the increasing expression of Cleaved caspase-3.And ursolic acid inhibited migration of Tca8113 cells by decreasing the expression of p-Fak and Fak.Conclusions Ursolic acid significantly inhibited the growth and migration of tongue cancer Tca8113 cells,which may be related to the regulation of Bcl-2/BAX apoptotic pathway and inhibition of Fak activation.