前列腺肿癌__(期刊论文)

前列腺特异性抗原启动子调控促凋亡基因caspase7载体的构建及其对前列腺癌细胞的特异性杀伤作用

http://www.100md.com 《第四军医大学学报》 2003年第8期

前列腺肿癌,,前列腺肿癌；启动区(遗传学)；脱噬作用；克隆，分子,0引言,1材料和方法,2结果

     (第四军医大学：西京医院泌尿外科，基础部生物化学与分子生物学教研室，陕西西安 710033)

    Overexpression of prostatespecific antigen promoterdriven Procaspase7 for the induction of therapeutic apoptosis in prostate cancer

    ZHANG Geng, WANG He, ZHANG Bo, YU CuiJuan, WU GuoJun, QIN WeiJun, MENG YanLing, WANG ChengJi, YANG AnGang

    Department of Urology Surgery, Xijing Hospital, Department of Biochemistry & Molecular Biology，Faculty of Preclinical Medicine, Fourth Military Medical University, Xian 710033, China

    【Abstract】AIM：To construct the eukaryotic expression vector that specifically drives pro-caspase-7 by prostate-specific antigen promoter. By employing the prostate-specific antigen promoter (PSAP), we investigated whether PSAPdriven procaspase7 could induce death of prostate cancer cells and whether the cytotoxicity was restricted to cells of prostate origin. METHODS: We extracted genomic DNA from benign prostate tissues and obtained prostate specific antigen promoter(PSAP)fragment by polymerasechainreaction (PCR).Using gene technology, we replaced the pCMV on pCDNA3 by PSAP and obtained PSAPpCDNA3 vector. Then the pCMV on pIRES2EGFP and pIRES2EGFPCsp7 was replaced by PSAP after the confirmation of its sequence and PSAPpIRES2EGFP and PSAPpIRES2EGFPCsp7 eukaryotic expression vectors were obtained respectively. Lipofectionmediated gene transfers were performed with the two obtained vectors and a control plasmid, pIRES2EGFP, in human prostate cancer cell line PC3 m and 2 other cell lines (Hep2 [human cancer of larynx] and MC3T3 [mice fibroblast]). Light and electron microscopy were used to observe the proliferation and apoptosis of all cell lines. A PSAPdriven EGFP was also used to estimate the expression of the PSAPdriven transcripts. Immunohistochemistry methods were used to evaluate the expression of caspase7 protein. RESULTS： All the constructed vectors were verified by enzyme digestion and DNA sequencing. After transfected with pIRES2EGFP, PC3 m, Hep2 and MC3T3 all showed green fluorescence. After transfected with PSAPpIRES2EGFP, only PC3m showed green fluorescence and no apoptosis cells appeared in all the three transfected groups. After transfected with PSAPcaspase7pIRES2EGFP, PC3 m showed the strong growth inhibition but the other two cell lines showed no changes. No caspase7 was detected in the Hep2 and MC3T3 cell lines after transfection. Morphologically, some of the PC3m transfected cells manifested apoptotic features. CONCLUSION: PSAPdriven procaspase7 gene therapy inhibits the growth of human prostate cancer cells and the cytotoxic effect is restricted to the cells of prostate origin. ......

百拇医药网 http://www.100md.com/html/DirDu/2006/02/19/87/28/34.htm

您现在查看是摘要页，全文长 15649 字符。