摘要
目的 研究扶正祛邪方对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响及其可能的机制.方法 扶正祛邪方作用于人卵巢癌SKOV3细胞后,通过MTT、细胞克隆、细胞划痕和Transwell实验,检测对SKOV3细胞增殖、凋亡、侵袭和迁移的影响;通过Wes-tern blotting和实时PCR检测负性表观遗传调控蛋白EZH2及其相关蛋白E-cadherin,以及细胞凋亡相关蛋白Bax、Bcl-2的表达水平.结果 扶正祛邪方可呈浓度及时间依赖性提高SKOV3细胞的生长抑制率,可明显抑制SKOV3细胞的增殖、侵袭和迁移能力;Western blotting及实时PCR结果表明,扶正祛邪方与GSK126联合能抑制EZH2及Bcl-2转录,促进Bax、E-cadherin转录,下调EZH2、Bcl-2蛋白表达,促进Bax、E-cadherin蛋白表达.结论 扶正祛邪方能抑制人卵巢癌SKOV3细胞增殖和侵袭,诱导其发生凋亡,可能是通过抑制EZH2参与调控E-cadherin表达及卵巢癌细胞的增殖、侵袭和迁移,并通过Bcl-2及Bax调控卵巢癌细胞的凋亡.
Abstract
Objective To investigate the effect of Fuzhengquxie prescription on the proliferation,apoptosis,invasion,and migration of ovarian cancer cells and its associated mechanism.Methods After Fuzhengquxie prescription was applied to human ovarian cancer SKOV3 cells,the effects on cell proliferation,apoptosis,invasion,and migration were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide,cell cloning,cell scratch,and Transwell assay experiments.Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blotting were used to determine the expression levels of the negative epigenetic regulatory protein,EZH2;its related protein,E-cadherin;and the apoptosis-related proteins,Bax and Bcl-2.Results Fuzhengquxie prescription inhibited the growth rate of SKOV3 cells in a concentration-and time-dependent manner,and significantly inhibited the proliferation,invasion,and migration of SKOV3 cells.Western blotting and qRT-PCR results showed that Fuzhengquxie prescription combined with GSK126 inhibited the transcription of EZH2and Bcl-2,promoted the transcription of Baxand E-cadherin,down-regulated the expression of EZH2 and Bcl-2 proteins,and promoted the expression of Bax and E-cadherin proteins.Conclusion Fuzhengquxie prescription inhibited the proliferation,invasion,and migration of SKOV3 cells and induced their apoptosis.It may be involved in regulating the E-cadherin-mediated proliferation,invasion,and migration of ovarian cancer cells by inhibiting the epigenetic regulatory protein EZH2,and regulating the apop-tosis of ovarian cancer cells mediated by Bcl-2 and Bax.
基金项目
国家自然科学基金(U1804181)
河南省中医药局科学研究专项(20-21ZY1036)
河南省科技攻关计划(182102311187)