摘要
目的:探讨核转录因子神经胶质瘤关联癌基因同源物1(glioma associated oncogene homolog 1,Gli-1)对转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人胃癌SGC-7901细胞发生上皮间质转化(epithelial-mesenchymal transition,EMT)的作用机制。方法:体外采用10 ng/ml TGF-β1对胃癌SGC-7901细胞进行处理,使用倒置显微镜观察细胞形态变化,RT-PCR和Western blot检测EMT上皮表型蛋白E-cadherin和间质表型蛋白Vimentin的表达水平; Transwell细胞侵袭实验检测细胞侵袭能力变化,检测TGF-β1对SGC-7901细胞发生EMT的影响;同时采用RT-PCR和Western blot检测Gli-1的mRNA和蛋白表达水平。随后进一步采用Gli-1基因特异性阻断剂GANT 61阻断Gli-1表达,并使用TGF-β1(10 ng/ml)对SGC-7901细胞进行处理,RT-PCR和Western blot检测Gli-1、E-cadherin和Vimentin mRNA及其蛋白表达水平的改变,并使用Transwell细胞侵袭实验检测阻断Gli-1对SGC-7901细胞侵袭能力的影响。结果:TGF-β1可以诱导人胃癌SGC-7901细胞发生上皮间质转化并促进细胞侵袭。TGF-β1可以在mRNA和蛋白水平下调上皮表型蛋白E-cadherin的表达、提高Gli-1和间质表型蛋白Vimentin的表达。TGF-β1可以明显提高SGC-7901细胞侵袭,而阻断Gli-1后可以抑制TGF-β1诱导的人胃癌SGC-7901细胞上皮间质转化和细胞侵袭。结论:胃癌SGC-7901细胞中Gli-1可能参与TGF-β1介导的EMT的发生,Gli-1可能作为胃癌基因治疗中的有效靶点发挥作用。
Objective： To investigate the role of Gli-1 in TGF-β1 induced epithelial-mesenchymal transition（ EMT） and invasion in gastric cancer cell line SGC-7901. Methods： Cultured SGC-7901 cells were treated with 10 ng/ml TGF-β1 for 24 h. The morphological changes were observed under phase-contrast microscopy. The mRNA and protein expression levels of Gli-1,EMT relative marker E-cadherin and Vimentin were detected by RT-PCR and Western blot. The effect of TGF-β1 on invasion ability of SGC-7901 cells were detected with Transwell method. Secondly,after co-treating with 10 ng/ml TGF-β1 and 10 μm GANT 61,RT-PCR and Western blot were used to check the mRNA and protein expression of Gli-1,E-cadherin and Vimentin. Transwell invasion assay was performed again to monitor cell invasion potentiality. Results： The results demonstrated that TGF-β1 could induce SGC-7901 cells EMT and enhance cell invasion. The mRNA and protein level of epithelial marker E-cadherin was downregulated by TGF-β1,whereas the mRNA and protein expression level of Gli-1 and mesenchymal maker protein Vimentin were elevated. Furthermore,TGF-β1 could significantly enhance the invasion ability of SGC-7901 cells,however,co-treated Gli-1 inhibitor GANT 61 with TGF-β1 significantly suppressed TGF-β1 induced EMT and cell invasion. Conclusion： Gli-1 participates in EMT induced by TGF-β1 in gastric cancer cell line SGC-7901.
作者
赵伟锋
陈雪姣
付蕾
胡金龙
卢创新
仓顺东
王朝杰
Zhao Weifeng;Chen Xuejiao;Fu Lei;Hu Jinlong;Lu Chuangxin;Cang Shundong;Wang Chaojie(Department of Medical Oncology,the People＇s Hospital of Henen Pros#tee,Henan Zhengzhou 450000,China)
出处
《现代肿瘤医学》
CAS
2018年第22期3530-3535,共6页
Journal of Modern Oncology
关键词
胃癌
转化生长因子-β1
核转录因子神经胶质瘤关联癌基因同源物1
上皮间质转化
侵袭
gastric cancer
transforming growth factor -β1 （ TGF -β1 ）
glioma associated oncogene homolog 1 （ Gli- 1 ）
epithelial - mesenchymal transition （EMT）
invasion
