紧密连接蛋白-6基因对结肠癌小鼠模型肿瘤生长影响及其表达水平  

Effects of claudin-6 gene on tumor growth in colon cancer mouse model and its expression level in tumor tissues

作　　者：唐小龙[1,2] 何一[1] 蒙长远 闫在华 李利发 吴溢[1] 侯松林[1,2] 周彤 Tang Xiaolong;He Yi;Meng Changyuan;Yan Zaihua;Li Lifa;Wu Yi;Hou Songlin;Zhou Tong(Department of Gastrointestinal Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;Institute of Hepatobiliary Pancreaticoenteropathy,North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院胃肠外科,南充637000 [2]川北医学院肝胆胰肠疾病研究所,南充637000

出　　处：《中华实验外科杂志》2020年第3期468-472,共5页Chinese Journal of Experimental Surgery

摘　　要：目的观察紧密连接蛋白(claudin)-6基因对结肠癌小鼠模型肿瘤生长的影响。方法将40只雄性C57BL/6小鼠(购自西安交通大学实验动物中心)分采用随机对照组原则分为对照组(10只)和模型组(30只),模型组通过腹腔注射氧化偶氮甲烷(AOM)的方式诱导结肠癌模型,将建模成功的小鼠随机分为两组,即模型组(10只)和claudin-6组(10只)。其中claudin-6尾静脉注射claudin-6过表达质粒,每周1次,连续4周。模型组注射等量的阴性对照(NC)质粒,对照组腹腔注射等剂量的生理盐水。干预后将小鼠拉颈处死取出结肠癌组织和正常结肠癌组织,分别检测瘤体的体积和质量。采用苏木精-伊红(HE)染色检测建立模型,采用免疫组织化学染色检测组织中claudin-6蛋白的表达,采用定量聚合酶链反应(qPCR)检测claudin-6 mRNA的水平,采用蛋白质印迹法(Western blot)检测claudin-6蛋白表达水平,采用微血管密度(MVD)检测结肠癌组织为血管生成。采用单因素方差分析(ANOVA)以评估组间差异,两两分析采用t检验。结果干预后claudin-6组的肿瘤体积和质量分别为(0.63±0.18)cm3和(0.57±0.15)g,均显著降低并低于对照组的(1.61±0.29)cm3和(1.46±0.34)g(t=6.754、8.695,P<0.05),差异有统计学意义。干预后模型组结肠癌组织中mRNA(1.98±0.21)和claudin-6蛋白(1.27±0.17)水平显著低于对照组(5.68±0.46和3.95±0.31,t=12.357、15.674,P<0.05),claudin-6组的mRNA(3.58±0.32)和claudin-6蛋白(2.46±0.27)水平显著高于模型组(1.98±0.21和1.27±0.17,t=5.647、6.358,P<0.05)。模型组的MVD水平(27.64±5.32)显著高于对照组(7.43±1.74,t=4.847,P<0.05),claudin-6组MVD水平(16.99±4.14)显著高于对照组(7.43±1.74,t=8.472,P<0.05),差异均有统计学意义。结论过表达claudin-6具有抑制结肠癌生长和血管生成的作用,提示claudin-6在结肠癌中的抑癌作用。Objective To investigate the effects of tight junction protein claudin-6 gene on tumor growth in colon cancer mouse models.Methods Forty male C57BL/6 mice were divided into the control group(n=10)and the model group(30)according to the principle of random control.The model group was induced into the colon cancer model by intra-abdominal injection of AOM.Claudin-6 overexpressed plasmid was injected into the mice according to the previously described method once a week for 4 weeks.The model group was injected with the same amount of NC plasmid,while the control group was intraperitoneal injected with the same amount of normal saline.After the intervention,the mice were killed by neck pulling to remove colon cancer tissue and normal colon cancer tissue,and the volume and mass of tumor body were detected respectively.HE staining was used to verify the established model,immunohistochemical staining was used to detect the expression of claudin-6 protein in the tissues,quantitative reverse transcriptase-polymerase chain reaction(qPCR)was used to detect the expression level of claudin-6 mRNA,Western blotting was used to detect the expression level of claudin-6 protein,and microvessel density(MVD)was used to detect the angiogenesis of colon cancer tissue.Results After intervention,the tumor volume[(0.63±0.18)cm3]and mass[(0.57±0.15)g]in the claude-6 group were significantly reduced as compared with those in the control group[(1.61±0.29)cm3 and(1.46±0.34)g](t=6.754,8.695,P<0.05).After intervention,mRNA(1.98±0.21)and claude-6 protein(1.27±0.17)levels in colon cancer tissues in the model group were significantly lower than those in the control group[(5.68±0.46)and(3.95±0.31,t=12.357,15.674,P<0.05].MRNA(3.58±0.32)and claudin-6 protein(2.46±0.27)levels in the claude-6 group were significantly higher than those in the model group(1.98±0.21,1.27±0.17,t=5.647,6.358,P<0.05).The MVD level in the model group(27.64±5.32)was significantly higher than that in the control group(7.43±1.74,t=4.847,P<0.05),and that in the cla

关 键 词：结肠癌 紧密连接 紧密连接蛋白-6 微血管 

分 类 号：R735.35[医药卫生—肿瘤] R-332[医药卫生—临床医学]