水苏碱调节Shh/Gli1信号通路对口腔癌细胞迁移、侵袭和上皮间质转化的影响

Effects of Stachydrine on the Migration,Invasion,and Epithelial-Mesenchymal Transition of Oral Cancer Cells by Regulating the Shh/Gli1 Signaling Pathway

目的探讨水苏碱(Sta)调节超音速刺猬因子(Shh)/胶质瘤相关癌基因同源物1(Gli1)信号通路对口腔癌(OC)细胞迁移、侵袭和上皮间质转化(EMT)的影响。方法采用蛋白质印迹技术检测Shh、Gli1在正常口腔上皮角质细胞HOK和OC细胞株系(Cal-27、Tca8113、SCC-9、HSC-3)中的表达,选择其中高表达的HSC-3细胞为研究对象,随机分为Control组、L-Sta组、M-Sta组、H-Sta组、PM组。EdU染色检测HSC-3细胞增殖;划痕实验检测HSC-3细胞的迁移;Transwell实验检测HSC-3细胞侵袭;采用蛋白质印迹技术检测Sta对E-cadherin、N-cadherin、Vimentin、Shh、Gli1蛋白表达的影响;小鼠荷瘤实验验证Sta对OC肿瘤生长和OC组织Shh、Gli1蛋白表达的影响。结果L-Sta组、M-Sta组、H-Sta组HSC-3细胞EdU阳性细胞率、划痕愈合率、侵袭数、N-cadherin、Vimentin、Shh、Gli1表达低于Control组,E-cadherin表达高于Control组(P<0.05);与H-Sta组相比,PM组EdU阳性细胞率、划痕愈合率、侵袭数、N-cadherin、Vimentin、Shh、Gli1表达升高,E-cadherin表达降低(P<0.05)。Sta组移植瘤质量、体积、Shh阳性率、Gli1阳性率低于对照组(P<0.05)。结论Sta可以抑制OC细胞迁移、侵袭和EMT,其机制可能是通过调节Shh/Gli1信号通路实现的。

Objective To investigate the effects of stachydrine(Sta)on the migration,invasion,and epithelial-mesenchymal transition(EMT)of oral cancer(OC)cells by regulating the sonic hedgehog(Shh)/glioma-associated oncogene homolog 1(Gli1)signaling pathway.Methods Western blot(WB)was used to detect the expression of Shh and Gli1 in normal human oral keratinocyte(HOK)and OC cell lines(Cal-27,Tca8113,SCC-9,and HSC-3).HSC-3 cells with high expression of Shh and Gli1 were regarded as the study subjects and randomly separated into Control group,L-Sta group,M-Sta group,H-Sta group,and PM group.EdU staining was applied to detect the proliferation of HSC-3 cells.Scratch experiment was applied to detect the migration of HSC-3 cells.Transwell experiment was applied to detect the invasion of HSC-3 cells.WB was applied to detect the effects of Sta on E-cadherin,N-cadherin,Vimentin,Shh,and Gli1 protein expression.Mouse tumor bearing experiment was applied to verify the effects of Sta on the growth of OC tumors and the expression of Shh and Gli1 proteins in OC tissues.Results The EdU positive cell rate,scratch healing rate,invasion number,N-cadherin,Vimentin,Shh,and Gli1 expression of HSC-3 cells in the L-Sta,M-Sta,and H-Sta groups were lower than those in the Control group,while the expression of E-cadherin was higher than that in the Control group(P<0.05).Compared with the H-Sta group,the PM group showed an increase in EdU positive cell rate,scratch healing rate,invasion number,N-cadherin,Vimentin,Shh,and Gli1 expression,and a decrease in E-cadherin expression(P<0.05).The quality,volume,Shh positive rate,and Gli1 positive rate of transplanted tumors in the Sta group were lower than those in the Control group(P<0.05).Conclusion Sta can inhibit the migration,invasion,and EMT of OC cells,which may be achieved by regulating the Shh/Gli1 signaling pathway.