阐明Wnt10A基因在胃癌细胞增殖及迁移中的功能及探索潜在的分子机制,为胃癌诊断、治疗提供新的靶点分子.以荧光定量PCR及免疫印迹技术检测基因表达,以RNAi技术敲减Wnt10A基因,以MTT法、划痕及transwell实验检测细胞行为学变化.结果显示,Wnt10A基因在胃癌组织中表达量均显著高于癌旁对照,平均差异达到3倍.并且Wnt10A基因在胃癌细胞系中表达量也高于正常细胞GES.当Wnt10A基因在胃癌细胞AGS中表达被成功下调约60%后,AGS细胞增殖率下降40%,迁移率下降约40%,侵袭能力也降低约70%.免疫印迹实验表明Wnt10A下调后,β-catenin、Cyclin D、TCF以及Myc基因表达量下调,而DKK1与GSK3β 表达增强,与LGK-974处理结果一致.Wnt10A基因通过模拟激活Wnt/β-catenin/Myc信号通路促进胃癌细胞增殖与迁移,发挥促癌基因的功能.%This work aims to elucidate the function of gene Wnt10A during the proliferation and migration of gastric cancer cells and the underlying molecular mechanism,for providing new target molecule in the diagnosis and treatment of gastric cancer. Real-time quantitative PCR and Western bolt analysis were used to detect gene expression ;RNAi technology was applied to knockdown and decrease the expression of Wnt10A,and MTT assay,wound healing assay and transwell assay were employed to detect the biological behavior of gastric cancer cells. Results showed that gene Wnt10A expressed more highly in tumor tissues than that in adjacent normal tissues,approximately 3 times. The level of Wnt10A in gastric cancer cell lines was also higher than the normal cells GES. The proliferation rate of AGS cells reduced by 40%,the migration rate decreased by 40%,and the invasive ability was also downregulated about 70%,when the expression level of gene Wnt10A in AGS cells decreased about 60%. Furthermore,Western blot analysis demonstrated that the expressions of β-catenin,Cyclin D,TCF and gene Myc reduced after Wnt10A knockdown in AGS cells,while the expression levels of DKK1 and GSK3β increased,which was in consistent with the results by the treatment of LGK-974,a specific inhibitor against Wnt/β-catenin signaling. Conclusively,gene Wnt10A promotes the proliferation and migration of gastric cancer cells through simulating activation of Wnt/β-catenin/Myc signaling pathway,i.e.,functions as cancer-promoting gene.
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