乳鼠结肠组织体外培养过程中TGF-β1对上皮细胞极性特征的影响  

TGF-β1 influences polarity characteristics of epithelial cells during process of culture of colon tissues from neonatal rats in vitro

作　　者：付道时[1] 文彬[1] 张凌杭 李茹柳[1] 

机构地区：[1]广州中医药大学脾胃研究所,广东省广州市510405

出　　处：《世界华人消化杂志》2014年第29期4386-4392,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81173257;中央财政支持地方高校发展专项资金"重大难治性脾胃病防治协同创新平台"基金资助项目;No.财教[2013]338号~~

摘　　要：目的:建立乳鼠结肠组织器官培养体系,分析转化生长因子β1(transforming growthfactor-β1,TGF-β1)对组织上皮细胞表面极性特征的影响.方法:在乳鼠结肠组织器官培养过程中添加10ng/mL TGF-β1干预48 h后,采用免疫组织化学、RT-PCR技术检测培养组(无添加TGF-β1)和干预组结肠组织中Crumbs3、E-cadherin、α-平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)的表达分布变化.结果:贴壁培养48 h后,可见培养组(无添加TGF-β1)组织块周围有铺路石样细胞爬满培养板,Crumbs3主要表达在上皮细胞顶端膜位置(P<0.01),E-cadherin主要表达在结肠组织黏膜上皮层,α-SMA沿隐窝轴单层表达和少量表达在平滑肌层.而添加TGF-β1干预后的结肠组织周边细胞形态大小不一,E-cadherin主要表达在细胞质,并呈低表达(P<0.05),α-SMA则大量表达在活化态的肌成纤维细胞和间充质中(P<0.01).培养组E-cadherin、Crumbs3的mRNA的表达量在48 h后明显低于干预组(P<0.05),而干预组中α-SMA的mRNA表达高于培养组(P<0.01).结论:TGF-β1可诱导乳鼠结肠上皮细胞极性蛋白Crumbs3的表达下降并导致肠上皮细胞表型的改变.AIM： To establish a neonatal rat colon tissue culture system and to analyze the effect of transforming growth factor-β1（TGF-β1） on polarity characteristics of epithelial cells.METHODS： Forty-eight hours after 10 ng/mL TGF-β1 was added during the process of culture of colon tissues from neonatal rats, the expression changes of Crumbs3, E-cadherin and alpha smoothmuscle actin（α-SMA） in the control group（without TGF-β1） and intervention group（with TGF-β1） were detected by IHC and RT-PCR.RESULTS： Pavement-like cells could be seen around tissues in the control group after 48 h; Crumbs3 was mainly located at the apical membrane of epithelial cells（P〈0.01）; a single layer of α-SMA-positive myofibroblasts along the crypt axis was visible, and α-SMA was lowly expressed in the smooth muscle. However, cells in the intervention group showed different morphology and size; E-cadherin was mainly present in the cytoplasm, and E-cadherin expression was lower than that in the control group（P〈0.05）; α-SMA was abundantly expressed in activated fibroblasts（P〈0.01）. The mRNA expression of E-cadherin and Crumbs3 in the control group was significantly lower than that in the intervention group（P〈0.05）, while the mRNA expression of α-SMA in the intervention group was significantly higher than that in the control group（P〈0.01）. CONCLUSION： TGF-β1 induced down-regulation of Crumbs3 and then dramatically changed the phenotype of epithelial cells.

关 键 词：转化生长因子Β1 器官培养 极性 细胞表型 肿瘤微环境 

分 类 号：R346[医药卫生—基础医学]