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

doi:10.3969/j.issn.2095-4344.2013.46.008

Abstract

Abstract:

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 urothelium cells and induce intestinal epithelial differentiation, thus inducing the development of cystitis glanduaris.

Key words: transcription factor, epithelial cells, chorionic villi, metaplasia

CLC Number:

R318

Lin Ming-en, Deng Bi-hua, Lü Yi-song, Rong Lu, Yao You-sheng. Construction of human caudal type homeobox transcription factor 2 retrovoral vector and transfection of urothelium cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(46): 8024-8029.

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