四君子汤含药血清通过抑制RhoA/Rac1/Cdc42通路影响卵巢癌细胞的上皮间质转化  

Sijunzi Decoction containing serum affects the epithelial mesenchymal transition of ovarian cancer cells by inhibiting the RhoA/Rac1/Cdc42 pathway

作　　者：李凡 张学林[1] 冯静茹[1] 郝洁[1] 周蕾 LI Fan;ZHANG Xuelin;FENG Jingru;HAO Jie;ZHOU Lei(the Second Hospital Affiliated to Hebei North University,Zhangjiakou 075100,China)

机构地区：[1]河北北方学院附属第二医院,张家口075100

出　　处：《中国比较医学杂志》2021年第8期29-37,共9页Chinese Journal of Comparative Medicine

基　　金：河北省医学科学研究课题计划项目(20200496)。

摘　　要：目的探索四君子汤含药血清对卵巢癌细胞增殖迁移、侵袭及上皮间质化的影响,并初步研究其作用机理。方法体外培养人卵巢癌细胞(SKOV3)和人卵巢上皮细胞,随机分为空白对照组、舒尼替尼组、四君子汤Ⅰ组、Ⅱ组和Ⅲ组。噻唑蓝法检测各组SKOV3细胞和人卵巢上皮细胞增殖变化情况;划痕实验和侵袭(transwell)实验检测各组SKOV3细胞迁移和侵袭能力;蛋白免疫印迹分析法检测各组SKOV3细胞上皮型钙粘蛋白(Ecadherin)、神经性钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)、Ras同源基因家族成员A(RhoA)、Ras相关的C3肉毒素底物1(Rac1)和细胞分裂周期蛋白42(Cdc42)蛋白表达情况;实时荧光定量PCR(qRT-PCR)法检测各组SKOV3细胞RhoA、Rac1和Cdc42 mRNA表达变化。结果各组人卵巢上皮细胞存活率差异无统计学意义(P>0.05)。与空白对照组相比,舒尼替尼组SKOV3细胞存活率、划痕愈合率、侵袭数量、N-cadherin、Vimentin蛋白、RhoA、Rac1、Cdc42 mRNA和蛋白表达显著降低(P<0.05),E-cadherin蛋白表达显著升高(P<0.05)。四君子汤Ⅰ、Ⅱ、Ⅲ组SKOV3细胞存活率、划痕愈合率、侵袭数量、N-cadherin、Vimentin蛋白、RhoA、Rac1、Cdc42 mRNA和蛋白表达依次降低(P<0.05),E-cadherin蛋白表达依次升高(P<0.05),呈剂量依赖性。四君子汤Ⅲ组上述指标和舒尼替尼组比较差异无统计学意义(P>0.05)。结论四君子汤含药血清能够抑制人卵巢癌细胞增殖、迁移、侵袭和上皮间质化,可能与RhoA/Rac1/Cdc42信号通路激活受阻相关。Objective To explore the effects of Sijunzi Decoction containing serum on the proliferation,migration,invasion,and epithelial mesenchymal transition of ovarian cancer cells,and the mechanism involved.Methods Human ovarian cancer cells(SKOV3)and human ovarian epithelial cells were cultured in vitro and randomly divided into blank control,sunitinib,and Sijunzi Decoction Ⅰ,Ⅱ,and Ⅲ groups.The methyl thiazolyl tetrazolium assay was used to detect the proliferation of SKOV3 cells and human ovarian epithelial cells.The migration and invasion abilities of SKOV3 cells were examined using the scratch test and transwell test.The protein expressions of E-cadherin,N-cadherin,vimentin,RAS homolog gene family member A(RhoA),Ras-related C3 botulinum toxin substrate 1(Rac1),and cell division cycle 42(Cdc42)in SKOV3 cells were detected by western blotting.In addition,the mRNA expressions of RhoA,Rac1,and Cdc42 in SKOV3 cells were detected by real-time quantitative PCR(qRT-PCR).Results There was no significant difference in the survival rate of human ovarian epithelial cells among the groups(P>0.05).Compared with the blank control group,the survival rate,wound healing rate,invasion number,protein expressions of N-cadherin and vimentin,and mRNA and protein expressions of RhoA,Rac1,and Cdc42 in SKOV3 cells in the sunitinib group were significantly lower(P<0.05),and the protein expression of E-cadherin was significantly higher(P<0.05).The survival rate,wound healing rate,invasion number,protein expressions of N-cadherin and vimentin,and mRNA and protein expressions of RhoA,Rac1,and Cdc42 in SKOV3 cells in the Sijunzi Decoction Ⅰ,Ⅱ,and Ⅲ groups were decreased(P<0.05),and the protein expression of E-cadherin was increased(P<0.05)dose-dependently.There was no significant difference in these indexes between the Sijunzi Decoction Ⅲ and sunitinib groups(P>0.05).Conclusions Sijunzi Decoction containing serum inhibits the proliferation,migration,invasion,and epithelial mesenchymal transition of human ovarian cancer cells,which m

关 键 词：四君子汤含药血清 人卵巢癌细胞 Ras同源基因家族成员A Ras相关的C3肉毒素底物1 细胞分裂周期蛋白42 上皮间质化 

分 类 号：R-33[医药卫生]