TCF7L2与WNT7b相互作用促进胃癌细胞增殖与转移  

Study on the interaction of TCF7L2 and WNT7b to promote the metastasis of gastric cancer cells

作　　者：杨留洋 高倩 程民 徐婷娟 胡世莲 沈国栋 Yang Liuyang;Gao Qian;Cheng Min(Dept of Geriatrics,Anhui Provincial Hospital Affiliaied to Anhui Medieal University,Hefei230001;Anhui Prowincial Key Laboratory of Tumor Immunotherapy and Nutrition Therapy,Hefei 230001;School of Life Sciences,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学附属省立医院老年医学科,合肥230001 [2]肿瘤免疫与营养治疗安徽省重点实验室,合肥230001 [3]安徽医科大学生命科学学院,合肥230032

出　　处：《安徽医科大学学报》2020年第11期1729-1734,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81802391);安徽省中央引导地方科技发展专项项目(编号:2019b12030026、2019b11030012)。

摘　　要：目的探讨胃癌细胞中受TCF7L2表达调控的WNT7b及它们对胃癌细胞增殖与转移的影响。方法qPCR与PCR芯片技术检测临床标本与胃癌细胞株中TCF7L2与WNT7b等基因的表达情况;使用200 ng/ml重组人WNT7b蛋白处理SGC7901与N87等胃癌细胞株,qPCR与Western blot法检测TCF7L2的表达,双荧光素酶报告基因检测系统检测β-catenin/TCF结合活性;为分析WNT7b对TCF7L2 mRNA稳定性的影响采用2μg/ml放线菌素D处理N87细胞。采用CCK-8试剂盒检测胃癌细胞的增殖能力,细胞划痕与Transwell实验分析胃癌细胞的迁移能力。结果分析SGC7901/TCF7L2、N87/shTCF7L2及其对照细胞株的PCR芯片数据,发现受TCF7L2表达影响最大的WNT配体是WNT7b。通过17例胃癌标本检测发现胃癌组织TCF7L2与WNT7b的转录水平均高于配对的癌旁正常组织,且二者表达呈正相关关系。使用WNT7b处理N87细胞24 h即可发现细胞TCF7L2转录水平升高(P<0.001),WNT7b同时增强了TCF7L2的mRNA稳定性(P<0.05),TCF7L2的蛋白水平以及β-catenin/TCF结合活性也随之增高(P<0.05)。通过细胞学实验显示WNT7b与TCF7L2能够增强胃癌细胞的增殖与迁移能力(P<0.05)。结论胃癌组织中TCF7L2与WNT7b表达水平高于正常对照组织;WNT7b与TCF7L2之间存在相互促进表达的作用,共同发挥对胃癌细胞增殖与迁移能力的调控。Objective To investigate the WNT ligands regulated by TCF7 L2 in gastric cancer cells and their functions on the proliferation and metastasis of gastric cancer cells.Methods qPCR and PCR array methods were used to detect the expression of TCF7 L2 and WNT7 b genes in gastric cancer cell lines and clinical specimens.200 ng/ml recombinant human WNT7 b protein was used to treat gastric cancer cell lines SGC7901 and N87.qPCR,Western blot and dual-luciferase reporter gene detection system were used to detect TCF7 L2 andβ-catenin/TCF binding activity,respectively.2μg/ml actinomycin D treatment was used to analyze the effect of WNT7 b on TCF7 L2 mRNA stability in N87 cells.CCK-8 kit,wound healing and transwell methods were used to analyze the abilities of proliferation and migration of gastric cancer cells.Results By analyzing PCR array data of SGC7901/TCF7 L2,N87/shTCF7 L2 and their corresponding control cell lines,it was found that WNT7 b was the most remarkable in the influenced WNT ligands by TCF7 L2 expression.The detection of 17 gastric cancer specimens showed that the transcription levels of TCF7 L2 and WNT7 b in gastric cancer tissues were both higher than those in normal adjacent tissues,and there should be a positive correlation between the expressions of TCF7 L2 and WNT7 b.TCF7 L2 transcription levels increased after the 24 h treatment of WNT7 b protein on N87 cells(P<0.001).Meanwhile,TCF7 L2 mRNA stability was enhanced(P<0.05),and TCF7 L2 protein levels and theβ-catenin/TCF binding activity were increased(P<0.05).Moreover,it was demonstrated that WNT7 b and TCF7 L2 enhanced the proliferation and migration abilities of gastric cancer cells(P<0.05)through the cytology experiments.Conclution The expression levels of TCF7 L2 and WNT7 b in gastric cancer tissues are higher than those in normal control.There is a mutual expression enhancement between WNT7 b and TCF7 L2,which plays a role in regulating the proliferation and migration of gastric cancer cells together.

关 键 词：胃癌 肿瘤转移 WNT信号通路 TCF7L2 WNT7b 

分 类 号：R735.2[医药卫生—肿瘤]