Construction Of Recombinant Adenovirus With RPB-DR Promoter And TK Gene And Study Of Its Inhibition Effect On Tumor Cells

PrefaceIt is the most important reason making hormone therapy be palliative but not curable for prostate cancer that most malignant cells growth will eventually become hormonal independent. Some studies have discovered that there are two kinds of malignant cells coexisting in prostate cancer; one expresses the AR, while another not. The former makes the activity of AR elevated by sensitization and grow hormonal-independently. The latter is the most important cause that makes the prognosis worse. A treatment aiming at the two kinds of cells is our purpose.Nowadays, there are many gene therapy strategies for prostate cancer. However, because of lacking specificity to prostate cancer cells, most of them are limited for clinical application.Now two prostatic specific promoters, PSA and rPB, which have both two AREs and an ARR, were studied more and more. But the two promoters have activity only in the prostate cancer cells that express AR. On the contrary, they have no activity in those do not express AR. At the same time, some study displayed that all prostate cancer cells had retinoic acid receptor. Therefore, the two AREs of rPB were replaced by three copies of RARE relatively and we constructed new promoter named rPB-DR which was effect on hormonal dependent and hormonal independent prostate cancer cells. We linked it to TK suicide gene mediated by adenovirus vector and study of its inhibition effect on prostate cancer cells.PART ONE:Construction of recombinant adenovirus with rPB-DR promoter and TK geneObjective:To construct replication-defective adenovirus which was recombinated with rPB-DR promoter and TK gene.Methods:The rPB-DR promoter was linked to pBluescript SK-TK and constructed pBluescript SK-rPB-DR-TK, then cut the purpose gene rPB-DR-TK by enzyme and subcloned it into the MCS(multiple cloning site) of pShuttle, and linearized the pShuttle-rPB-DR-TK...