下调Lgr5表达对结肠癌细胞上皮-间质转化及血管生成相关因子的影响  

Effect of down-regulation of Lgr5 expression on epithelial-mesenchymal transition and angiogenesis related factors in colon cancer cells

作　　者：路明[1] 陈哲 LU Ming;CHEN Zhe(*Department of Gastroenterology,Jilin Central Hospital,Jilin,Jilin 132001,China)

机构地区：[1]吉林市中心医院胃肠外科,吉林吉林132001 [2]吉林市人民医院口腔科

出　　处：《青岛医药卫生》2021年第5期321-326,共6页Qingdao Medical Journal

摘　　要：目的观察下调G-蛋白偶联受体5(Lgr5)基因表达对结肠癌(Colon cancer,CC)细胞侵袭和迁移及上皮-间质转化(Epithelial-mesenchymal transition,EMT)和血管生成相关因子的影响。方法检测正常肠黏膜组织与结肠癌组织中Lgr5的表达差异;以正常肠黏膜细胞NCM460作为对照组,采用RT-PCR法和Western blot法从多种CC细胞株(HT-29、SW620、LoVo)中筛选出Lgr5相对表达量最高的肿瘤细胞株;下调LoVo细胞中Lgr5表达,将实验分为对照组(只转染Lip 2000)、si-NC组(转染Lip2000和无关序列)和si-Lgr5组(转染Lip2000和si-Lgr5序列),Transwell小室检测细胞侵袭能力,划痕实验检测细胞迁移能力;Western Blot法检测EMT相关蛋白波形蛋白(Vimentin)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)表达情况;酶联免疫吸附(ELISA)法检测血管内皮生长因子(Vascular endothelial growth factor,VEGF)和碱性成纤维细胞生长因子(Basic fibroblast growth factor,BFGF)的含量。结果与正常肠黏膜组织相比,Lgr5在结肠癌组织中表达升高(P<0.001);LoVo细胞中Lgr5表达量较NCM460细胞和其他CC细胞株(HT-29,SW620)高;与对照组比较,si-Lgr5组细胞侵袭能力和迁移能力被抑制(P<0.05),N-cadherin、Vimentin蛋白表达降低,E-cadherin表达升高,VEGF和bFGF含量降低(P<0.05)。结论下调Lgr5表达,能够抑制LoVo细胞侵袭、迁移,其机制可能与抑制EMT和血管生成有关。Objective To observe the effect of down-regulation of G-protein coupled receptor 5(Lgr5)gene expression on colon cancer(CC)cell invasion and migration,epithelial-mesenchymal transition(EMT)and angiogenesis-related factors.Methods The difference in the expression of Lgr5 between normal intestinal mucosal tissue and colon cancer tissue was detected.With normal intestinal mucosal cells NCM460 as control group,RT-PCR and Western blot methods were used to select the tumor cell line with the highest relative expression of Lgr5 from a variety of CC cell lines(HT-29,SW620,LoVo).The expression of Lgr5 in LoVo cells was down-regulated and the experiment was divided into control group(transfected with Lip 2000 only),si-NC group(transfected with Lip2000 and irrelevant sequences)and si-Lgr5 group(transfected with Lip2000 and si-Lgr5 sequences).Transwell chamber was used to detect cell invasion ability and scratch test was used to detect cell migration ability.Western Blot method was used to detect EMT-related protein Vimentin,E-cadherin,N-cadherin expression.Enzyme-linked immunosorbent(ELISA)method was used to detect vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)content.Results Compared with normal intestinal mucosal tissues,the expression of Lgr5 in colon cancer tissues was increased(P<0.001).The expression of Lgr5 in LoVo cells was higher than that in NCM460 cells and other CC cell lines(HT-29,SW620).Compared with control group,cell invasion and migration ability of si-Lgr5 group were inhibited(P<0.05),N-cadherin,Vimentin protein expression decreased,E-cadherin expression increased,VEGF and bFGF content decreased(P<0.05).Conclusion Down-regulating the expression of Lgr5 could inhibit the invasion and migration of LoVo cells and the mechanism might be related to the inhibition of EMT and angiogenesis.

关 键 词：结肠癌 G-蛋白偶联受体5 上皮间质转化 侵袭迁移 血管生成 

分 类 号：R735.35[医药卫生—肿瘤]