Cdx2反转录病毒载体的构建及转染膀胱上皮细胞的研究  

Construction of human caudal type homeobox transcription factor 2 retrovoral vector and transfection of urothelium cells

作　　者：林明恩[1] 邓毕华[2] 吕夷松[2] 荣禄[2] 姚友生[2] 

机构地区：[1]汕头大学医学院第一附属医院泌尿外科,广东省汕头市515041 [2]中山大学附属第二医院泌尿外科,广东省广州市510120

出　　处：《中国组织工程研究》2013年第46期8024-8029,共6页Chinese Journal of Tissue Engineering Research

基　　金：广东省医学科研基金立项课题(B2013267)~~

摘　　要：背景:尾侧型同源转录因子2在消化道尤其是小肠与结肠上皮的发育中起到关键作用。目的:构建尾侧型同源转录因子2反转录病毒表达载体pLNCX2-Cdx2,观察pLNCX2-Cdx2体外转染对人膀胱上皮细胞发生肠上皮化生的作用。方法:应用基因重组技术将人尾侧型同源转录因子2cDNA克隆到反转录病毒载体pLXSN2,酶切鉴定,包装到PA317细胞中。转染人膀胱尿路上皮细胞,应用荧光PCR及Western blot检测尾侧型同源转录因子2以及绒毛蛋白、LI-cadherin的表达情况。结果与结论:pLNCX2-Cdx2成功构建;细胞转染后尾侧型同源转录因子2的mRNA及蛋白水平表达明显增加;尾侧型同源转录因子2的过表达明显激活了绒毛蛋白与LI-cadherin的表达;转染后细胞可出现肠细胞样特征改变。结果提示体外尾侧型同源转录因子2过表达可使膀胱上皮细胞发生尾侧型同源转录因子2激活并产生肠上皮分化,从而诱导腺性膀胱炎发生。BACKGROUND： Caudal type homeobox transcription factor 2 plays an important role in the development of epithelium in digestive tract, especially the small intestine and colon. OBJECTIVE： To construct the retroviral expression vector of pLNCX2-caudal type homeobox transcription factor 2, and to observe the effect of in vitro transfection of pLNCX2-caudal type homeobox transcription factor 2 in intestinal metaplasia. METHODS： Gene recombinant technology was employed to clone human caudal type homeobox transcription factor 2 gene to the retroviral expression vector of pLNCX2, then identified with enzyme digestion and sequencing and packed to the PA317 cells. The plasmids were transfect in urothelium cells, real-time PCR and western blot were used to detect the expressions of caudal type homeobox transcription factor 2, villin and liverintestin- cadherin at the protein and mRNA levels. RESULTS AND CONCLUSION： The retroviral vector pLNCX2-caudal type homeobox transcription factor 2 was successfully constructed. The levels of caudal type homeobox transcription factor 2 protein and mRNA expressions in bladder urothelium cell transfected with pLNCX2-caudal type homeobox transcription factor 2 were higher than that in control. And the over-expression of caudal type homeobox transcription factor 2 could up-regulate the levels of villin and liverintestin-cadherin expressions. Interestingly, specific changes of intestine-like cells were seen in the bladder urothelium cells transfected by pLNCX2-caudal type homeobox transcription factor 2. Over-expression of caudal type homeobox transcription factor 2 can activate the caudal type homeobox transcription factor 2 of urethelium cells and induce intestinal epithelial differentiation, thus inducing the development of cystitis glanduaris.

关 键 词：组织构建 组织构建细胞学实验 尾侧型同源转录因子2 绒毛蛋白 肝肠钙粘连蛋白 肠上皮化生 膀胱上皮细胞 LI-CADHERIN 尿路上皮 腺性膀胱炎 省级基金 

分 类 号：R318[医药卫生—生物医学工程]