扶正抗癌汤含药血清调节EMT进程抑制人卵巢癌HO-8910PM细胞转移及侵袭作用的研究

Inhibitory effects of Fuzheng Kang-Ai decoction on migration and invasion of human ovarian carcinoma HO-8910PM cells by regulating the epithelial-mesenchymal transition process

目的研究扶正抗癌汤含药血清抑制HO-8910PM细胞转移及侵袭的作用及潜在机制。方法将48只SD大鼠随机分为正常组及扶正抗癌汤低(4.725 g/(kg·d))、中(9.45 g/(kg·d))、高(18.9 g/(kg·d))剂量组,灌胃给药,每天1次,共持续7 d;分离含药血清,用于孵育HO-8910PM细胞。通过划痕实验、Transwell实验、ELISA实验、实时定量PCR实验及Western blot实验检测相关指标变化情况。结果与正常组比较,低、中、高剂量扶正抗癌汤组含药血清处理HO-8910PM细胞24 h后,迁移率及穿膜细胞数均明显降低(P<0.01);与正常组比较,低、中、高剂量扶正抗癌汤组含药血清处理HO-8910PM细胞12 h、24 h和48 h后,FN蛋白表达明显降低(P<0.05或P<0.01);与正常组比较,低、中、高剂量扶正抗癌汤组含药血清处理HO-8910PM细胞24 h后,E-cadherin mRNA表达增加(P<0.01),而vimentin、N-cadherin m RNA表达及TGF-β1、p-Smad3蛋白表达显著减少(P<0.01)。结论扶正抗癌汤具有抑制人卵巢癌HO-8910PM细胞转移及侵袭的作用,该作用与阻碍TGF-β1/Smad3通路活化,进而调节EMT进程有关。

Objective To study the inhibitory effects of Fuzheng Kang-Ai(FZKA)decoction on the migration and invasion of human ovarian carcinoma(HO-8910 PM)cells and the underlying mechanisms.Methods Forty-eight SD rats were randomly divided into a control group,low-dose FZKA group(4.725 g/(kg·d)),medium-dose FZKA group(9.45 g/(kg·d)),and high-dose FZKA group(18.9 g/(kg·d)).The drug was administered by gavage once per day for 7 days.The decoction was isolated and used to incubate HO-8910 PM cells,and changes in the related indices were detected by the cell scratch test,transwell assay,ELISA,real-time quantitative PCR,and Western blot.Results Compared with the control group,after 24 hours of low-,medium-,and high-dose FZKA decoction treatment,HO-8910 PM cell mobility and invasion were significantly lower(P<0.01).Compared with the control group,after 12-,24-,and 48 h of low-,medium-,and high-dose FZKA decoction treatment of HO-8910 PM cells,FN protein expression decreased significantly(P<0.05 or P<0.01).Compared with the control group,after 24 h of low-,medium-and high-dose FZKA decoction treatment of HO-8910 PM cells,E-cadherin m RNA expression increased(P<0.01),while vimentin,N-cadherin mRNA,TGF-β1,and p-Smad3 expression decreased significantly(P<0.01).Conclusions FZKA can inhibit the migration and invasion of HO-8910 PM cells,by inhibiting TGF-β1/Smad3 pathway activation and further regulation of the EMT process.