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Cloning And Characterization Of The Promoter Of The Human Prostate Cancer Related New Gene PC-1

Posted on:2005-01-27Degree:DoctorType:Dissertation
Country:ChinaCandidate:J WangFull Text:PDF
GTID:1104360122498581Subject:Genetics
Abstract/Summary:PDF Full Text Request
The PC-1 was firstly identified and cloned by Zhou Jian-Guang et al. The expression of this gene is up-regulated in the advanced prostate cancer cell line C4-2, and the distribution of PC-1 in human organ is highly prostate specific. We also know that the expression of PC-1 is stimulated by androgen. Until now we have known little about its promoter which makes the most important role on the gene transcription. So in this paper we described the cloning and characterization of the PC-1 gene promoter.Firstly we identified a 5-kb genomic DNA fragment located in the 5' upstream of the PC-land cloned it into the expression vector pGL3-basic, at the same time we also cloned a series of 5' flanking deletion fragments into the expression vector. The transient transfection assays demonstrated that the 5-kb cloned promoter has the highest transcriptional activity among a series of 5' flanking deletion fragments, and even higher than the 6-kb promoter of the prostate specific antigen in the androgen ablation medium. We also found that the 5-kb promoter of PC-1 has the prostate cell-specific activity though we can detect its activity in the MCF-7 and 293 cells. The fragment between 4-kb and 5-kb from the translational start site of the PC-1 achieves the prostate-specific regulation activity and also displays the properties of the enhancer.Then we found an androgen receptor response element which located between 345 and 359 position from the translation start site of PC-1.The 5-kb promoter lost its regulation ability by androgen when we deleted the putative androgen receptor response element from this promoter.We also found that p53 repressed the transactivation of the PC-1 promoter. In the prostate cancer cell LNCaP we found that the repression of the transactivation partly achieves through the androgen receptor response element on the PC-1 promoter, while this ability lost in another prostate cancer cell C4-2. Maybe this is just one of the reasons that lead the higher expression level of PC-1 in C4-2.The prostate cell-specific transcription regulation ability of 5-kb PC-1 promoter andits higher transactivation make it to be suitable candidate for driving prostate-specific expression of therapeutic genes. The PC-1 5-kb promoter regulated the expression of p53 in the LNCaP cell and effectively repressed its growth while not affect the Hela cell which don't express the PC-1. We also found p53 expressed in C4-2B and 293 cells and induced the apoptotic features in these cells. All these results provide the basis for the application of the 5-kb PC-1 promoter.
Keywords/Search Tags:PC-1, promoter, prostate cancer, androgen receptor response element, p53
PDF Full Text Request
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