神经胶质成熟因子γ对宫颈癌细胞侵袭迁移的影响及其机制

Effect of glia maturation factor-γon invasion and migration of cervical cancer cells and its mechanism

目的探讨神经胶质成熟因子γ(glia maturation factor-γ,GMFG)在不同宫颈癌细胞中的表达情况及其对宫颈癌细胞上皮间质转化(epithelial-mesenchymal transition,EMT)的调控作用。方法利用Western blot检测人宫颈永生化鳞状细胞系Ect1/E6E7、不同宫颈癌细胞系SiHa、C33A、HeLa中GMFG的表达情况;选择SiHa细胞系为研究对象,利用慢病毒转染构建低表达GMFG的SiHa细胞系,即LV-GMFG-shRNA组,未处理的SiHa细胞为control组;采用细胞划痕实验和Transwell实验来研究48 h后GMFG对SiHa细胞迁移和侵袭的影响;利用Western blot检测SiHa细胞中GMFG低表达对EMT相关蛋白E-cadherin、N-cadherin、vimentin、Snail的影响。结果Ect1/E6E7细胞中几乎不表达GMFG;与C33A、HeLa细胞系相比,SiHa细胞系中GMFG的表达最丰富(P<0.05)。细胞划痕实验和Transwell检测结果均表明在48 h时,LV-GMFG-shRNA组细胞迁移及细胞侵袭均明显低于control组(P<0.05)。Western blot结果显示,与control组相比,LV-GMFG-shRNA组E-cadherin的表达水平升高,N-cadherin、Snail及vimentin的表达均下降(P<0.05)。结论GMFG在宫颈鳞癌细胞SiHa中高表达,且抑制GMFG的表达能降低宫颈鳞癌细胞的侵袭转移能力及EMT。

Objective To study the expression of glia maturation factor-γ(GMFG)in different cervical cancer cells and its regulation role in the epithelial-mesenchymal transition(EMT)of cervical cancer cells.Methods The expression of GMFG was detected in human cervical immortem squamous cell line Ect1/E6E7 and different cervical cancer cell lines SiHa,C33A and HeLa by Western blot.SiHa cells with low GMFG expression were successfully constructed by lentivirus transfection,namely LV-GMFG-shRNA group.Untreated SiHa cells were chosen as control group.Cell scratch assay and Transwell assay were used to assess the effect of GMFG on the migration and invasion of SiHa cells at 48 h.Western blot was used to detect the effect of low GMFG expression on EMT-related proteins including E-cadherin,N-cadherin,vimentin and Snail in SiHa cells.Results GMFG was hardly expressed in Ect1/E6E7 cells,and the expression of GMFG was higher in SiHa cells than that in C33A and HeLa cells(P<0.05).The abilities of migration and invasion in LV-GMFG-shRNA group were significantly lower than those in control group at 48 h by cell striation test and Transwell test(P<0.05).Compared to control group,the expression of E-cadherin was increased in LV-GMFG-shRNA group by Western blot(P<0.05),while the expression levels of N-cadherin,Snail and vimentin were decreased(P<0.05).Conclusion GMFG is highly expressed in squamous cell carcinoma of the cervix,and the inhibition of GMFG expression can reduce the abilities of invasion and metastasis and EMT of SiHa cells.