[Constructing recombinant plasmid with prostate-specific membrane antigen promoter and reverse enhancer and comparing the transcription activity of enhancers in different directions].
OBJECTIVE
To assess the specificity of prostate-specific membrane antigen(PSMA) promoter and enhancer in controlling gene expression and to compare the activity of enhancers in different directions for choosing the most suitable prostate-specific PSMA controlling elements.


METHODS
PSMA enhancer gene was amplified with PCR, then the enhancer gene was subcloned into the expressing vector pEGFP-PSMA(Pro) reversely to construct the recombinant plasmid pEGFP-PSMA(E(r)-p), which was transfected into different cell lines such as LNCaP, PC-3,MCF-7,A549. Green fluorescent protein (GFP) expression was observed and compared to other recombinants constructed previously.


RESULTS
The recombinant plasmid with reverse enhancer was successfully constructed, the PSMA promoter and enhancer showed modulating activity in PSMA-expressed cell line uniquely. PSMA enhancer could increase 30-fold transcriptional activity over the basal level achieved by PSMA promoter alone, and no impact of the direction on the activity of enhancer was noted.


CONCLUSIONS
PSMA promoter/ enhancer is specific to PSMA-expressed cells. The transcriptional activity of reverse enhancer is similar to that of enhancer. PSMA promoter/enhancer has the potential for use in targeted gene therapy of prostate adenocarcinoma.
Zeng
Hao
H
Department of Urology, West China Hospital of Sichuan University, Chengdu 610041, China.
Liao
Yong-Chuan
YC
Wang
Fang
F
Wu
Qi
Q
Yang
Yu-Ru
YR
D000230:Adenocarcinoma / Q000628:therapy; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011467:Prostate / Q000276:immunology / Q000378:metabolism; D017430:Prostate-Specific Antigen / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000628:therapy; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D014162:Transfection
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D017430:Prostate-Specific Antigen