【摘要】 目的:构建尾侧型同源转录因子2(Cdx2)逆转录病毒表达载体(pLNCX2-Cdx2);研究pLNCX2-Cdx2体内转染对大鼠膀胱上皮细胞的生物学作用。方法:用基因重组技术将Cdx2克隆到逆转录病毒载体(pLXSN2),酶切鉴定,并包装到PT67细胞中。通过膀胱灌注法转染大鼠膀胱;实验分pLNCX2-Cdx2、空载体及对照组;用实时定量聚合酶链反应、Western blot、MTT、苏木精-伊红染色及透射电镜分别验证转染效果。结果:酶切和测序表明成功构建pLNCX2-Cdx2;pLNCX2-Cdx2组Cdx2表达明显升高,并激活绒毛蛋白与肝肠钙粘连蛋白表达,高于另两组(P<0.05);膀胱上皮可见肠上皮细胞样改变。结论:体内Cdx2过表达可使大鼠膀胱上皮细胞Cdx2激活并产生肠上皮分化。更多还原

【Abstract】 Objective:To construct the retroviral expression vector of pLNCX2 caudal-related homeoboxgene(Cdx)2, identify the expression,transfect into rat bladder transitional epithelium,and to observe the expression of the targeted gene. Methods:Gene recombinant technology was employed to clone Cdx2 gene to the retroviral expression vector of pLNCX2, then to identify with enzyme digestion and sequencing. The plasmids were transfect in rat bladder. The successful gene transfer was confirmed by real-time PCR, Western blot, micro-scopy analyses and transmission electron microscopy.Results:The retroviral vector pLNCX2-Cdx2 was successfully constructed. The levels of Cdx2 protein and mRNA in bladder urothelium transfected with pLNCX2-Cdx2 were higher than those in control. The overexpression of Cdx2 up-regulated the levels of villin and liverintestin-cadherin and also raised the cell proliferation. The specific changes of intestine-like cell were seen in the urothelium cells. Conclusion:Overexpression of Cdx2 in vivo into vesical urothelium can up-regulate Cdx2 expression and activate the expression of villin and Ll-cadherin,and induces development of cystitis glanduaris.更多还原