穿心莲内酯抑制宫颈癌Hela细胞生长和迁徙的研究

Study on Inhibition of Andrographolide on the Growth and Migration of Cervical Cancer Cells

目的:研究穿心莲内酯对宫颈癌细胞株(Hela细胞)生长及迁徙的影响,并探究其相关作用机制。方法:体外培养宫颈癌Hela细胞,采用不同浓度穿心莲内酯(0,15,30,60μmol·L^(-1))处理细胞,穿心莲内酯浓度为0μmol·L^(-1)时为对照组。采用MTT法检测细胞增殖情况,Transwell小室检测细胞迁移能力变化,流式细胞术检测细胞凋亡水平变化,平板克隆实验检测细胞克隆形成情况,蛋白质免疫印迹实验检测内质网应激相关蛋白CHOP、凋亡相关蛋白前体caspase-3和caspase-9表达变化。结果:随着穿心莲内酯浓度的升高,Hela细胞存活率、迁移数及克隆形成率逐渐降低,凋亡率逐渐升高,与对照组比较差异均有统计学意义(P<0.05或P<0.01)。随着穿心莲内酯浓度的增加,Hela细胞中CHOP、钙黏附蛋白-E(E-cadherin)表达量逐渐升高,caspase-3、caspase-9、血管内皮生长因子(VEGF)及Ras相关的C3肉毒素底物1(Rac1)蛋白表达量逐渐降低(P<0.05)。结论:穿心莲内酯能够有效抑制Hela细胞生长及迁徙,其抑制迁徙的机制可能与提升E-cadherin蛋白表达及抑制VEGF、Rac1蛋白表达相关。

Objective: To study the effects of andrographolide on the growth and migration of cervical cancer cell lines( Hela cells),and explore its related mechanism. Methods: Cervical cancer Hela cells were cultured in vitro and were treated with different concentrations of andrographolide( 0,15,30 and 60 μmol·L^-1). The andrographolide concentration of 0 μmol·L^-1 was taken as the control group. The cell proliferation was detected by MTT assay,the cell migration ability was detected by Transwell chamber,the cell apoptosis was detected by flow cytometry,the cell clone formation was detected by plate clone assay,the expression levels of endoplasmic reticulum stress-related protein C/EBP-homologous protein( CHOP) and apoptosis-related protein precursors of caspase-3 and caspase-9 were detected by Western blot. Results: With the increase of andrographolide concentration,the survival rate,migration number and colony formation rate of Hela cells were decreased gradually,the apoptotic rate increased gradually,and there were significant differences between andrographolide groups and the control group( P < 0. 05 or P < 0. 01). With the increase of andrographolide concentration,the expression level of CHOP and E-cadherin protein in Hela cells were increased gradually while the expression levels of caspase-3,caspase-9,vascular endothelial growth factor( VEGF) and Ras-related C3 botulinumtoxinsubstrate1( Rac1) protein were decreased gradually( P < 0. 05). Conclusion: Andrographolide can effectively inhibit the growth and migration of Hela cells,and its mechanism of migration inhibition may be related to the promotion of E-cadherin protein expression and the inhibition of VEGF and Rac1 protein expressions.