摘要
目的 探讨Nanog与KDM5B对胃癌细胞迁移和侵袭能力的影响及分子调控机制。方法利用TGF-β1诱导人胃腺癌MGC-803细胞,并检测诱导后的MGC-803细胞中E-cadherin、N-cadherin、Nanog与KDM5B的表达水平;利用划痕实验、Transwell侵袭实验和免疫细胞化学实验检测过表达或敲低Nanog对MGC-803细胞迁移、侵袭、E-cadherin和N-cadherin表达水平的影响;用RT-qPCR实验和免疫细胞化学实验检测过表达或敲低Nanog对KDM5B表达水平的影响;最后,用Transwell侵袭实验和免疫细胞化学实验检测敲低KDM5B对MGC-803细胞侵袭能力和E-cadherin和N-cadherin表达水平的影响。结果 TGF-β诱导的MGC-803细胞中E-cadherin表达降低,而N-cadherin、Nanog与KDM5B的表达升高;过表达Nanog促进MGC-803细胞迁移、侵袭和N-cadherin、KDM5B的表达,抑制E-cadherin的表达,敲低Nanog得到相反的结果;敲低KDM5B后的MGC-803细胞侵袭能力和N-cadherin表达水平降低,E-cadherin表达水平增高。结论 Nanog正向调控KDM5B促进胃腺癌细胞迁移、侵袭和上皮间质转化。
Objective To explore the effect of Nanog and KDM5 B on migratory and invasive activities of gastric cancer(GC)cells and the molecular regulative mechanism involved in GC development.Methods Human gastric adenocarcinoma MGC-803 cells were treated with TGF-β1 and the expression levels of E-cadherin,N-cadherin,Nanog and KDM5 B in the induced cells were detected;The effect of Nanog overexpression or knockdown on cell migratory and invasive abilities,as well as the expressions of E-cadherin and N-cadherin,in MGC-803 cells were detected by wound healing assay,Transwell invasion assay and immunochemistry cellular(ICC) assay;The effect of Nanog on KDM5 B in MGC-803 cells was examined by RT-qPCR and ICC assays;Finally,the effect of KDM5 B knockdown on cell invasive activity and expression levels of E-cadherin and N-cadherin in MGC-803 cells were evaluated by Transwell invasion assay and ICC assay.Results The expression of E-cadherin was decreased,while the expressions of N-cadherin,Nanog and KDM5 B were increased in TGF-β1 induced MGC-803 cells.Nanog overexpression enhanced cell migratory and invasive abilities of MGC-803 cells,as well as the expressions of N-cadherin and KDM5 B,and inhibited the expression of E-cadherin.Nanog knockdown led to the opposite results;The invasive ability of MGC-803 cells and the expression level of N-cadherin were decreased,while the expression level of E-cadherin was increased according to KDM5 B knockdown.Conclusion Nanog positively regulates KDM5 B to promote gastric adenocarcinoma cell migration and invasion.
作者
刘艳坤
卢琳
刘晓辉
马丽丽
李玉凤
LIU Yankun;LU Lin;LIU Xiaohui;MA Lili;LI Yufeng(Central Laboratory,Tangshan People’s Hospital,Tangshan 063001,China)
出处
《中国煤炭工业医学杂志》
2022年第3期231-236,共6页
Chinese Journal of Coal Industry Medicine
基金
河北省自然科学基金精准医学联合基金培育项目(编号:H2021105019)。
