FKBP10促进胃癌细胞增殖与转移的机制研究

Mechanism of FKBP10 promoting proliferation and migration of gastric cancer cells

目的探讨FKBP10在胃癌进展中发挥的调控作用及其相关机制。方法利用生物信息学分析FKBP10在胃癌组织中的表达及其与胃癌患者预后的关系;利用qRT-PCR及Western blot技术检测胃癌细胞系及胃粘膜上皮细胞GES中FKBP10的mRNA及蛋白表达水平;平板克隆实验及Transwell实验检测FKBP10下调对胃癌细胞增殖及转移的影响;利用KEGG信号通路富集分析FKBP10参与的信号通路调控并通过Western blot技术进行验证。结果与正常胃黏膜上皮细胞GES相比,FKBP10在胃癌细胞系中的表达水平相对较高;利用siRNA技术降低FKBP10的表达后,胃癌细胞的增殖及转移能力相比对照组明显减弱;KEGG信号通路富集分析发现FKBP10可能参与调控上皮-间质转化进程,而TGF-β在该进程中发挥重要作用,Western blot结果表明FKBP10表达下调后上皮相关标记物表达水平上升而间质相关标记物表达下降,同时p-SMAD2/3表达水平亦随之下降。结论FKBP10可能通过调控EMT进程及TGF-β/SMAD信号通路,从而促进胃癌增殖与转移。

Objective To investigate the regulative role of FKBP10 in the progression of gastric cancer and the underlying mechanism.Methods Bioinformatics analysis was applied to detect the expression of FKBP10 in gastric cancer samples and normal samples,as well as analyze its relationship with prognosis.qRT-PCR and Western blotting were employed to measure its expression at mRNA and protein levels in gastric cancer cell lines BGC823,MGC803,SGC7901,MKN45 and AGS and human gastric epithelial cell line GES.Then colony formation and transwell assay were conducted to detect the effect of its down-regulation on the proliferation and migration of the cells.Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was used to reveal the FKBP10 involved signaling pathways,which was verified by Western blotting.Results FKBP10 was up-regulated in gastric cancer cell lines than the GES cells.Knockdown of FKBP10 by siRNA restricted the proliferative and migrative abilities of gastric cancer cells.KEGG analysis indicated that FKBP10 may be involved in the process of epithelial-mesenchymal transition(EMT)and TGF-βplayed important role in the process.Western blotting showed that down-regulation of FKBP10 resulted in increased expression of epithelial related biomarkers while decreased expression of mesenchymal related biomarkers,then the expression of p-SMAD2/3 were reduced simultaneously.Conclusion FKBP10 promotes the proliferation and migration of gastric cancer via regulating EMT and TGF-β/SMAD signaling pathway.