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

Lin Ming-en; Deng Bi-hua; L Yi-song; Rong Lu; Yao You-sheng

doi:10.3969/j.issn.2095-4344.2013.46.008

Abstract：BACKGROUND:Caudal type homeobox transcription factor 2 plays an important role in the development of epithelium in digestive tract, especial y the smal intestine and colon. <br> 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. <br> 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, vil in and liverintestin-cadherin at the protein and mRNA levels. <br> RESULTS AND CONCLUSION:The retroviral vector pLNCX2-caudal type homeobox transcription factor 2 was successful y constructed. The levels of caudal type homeobox transcription factor 2 protein and mRNA expressions in bladder urothelium celltransfected 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 vil in 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 urothelium cells and induce intestinal epithelial differentiation, thus inducing the development of cystitis glanduaris.