当前位置: 首页 > 期刊 > 《第四军医大学学报》 > 2005年第12期
编号:11007068
PSMAIRESFCY1TK真核表达载体的构建及在前列腺癌细胞中的表达
关键词:前列腺肿瘤,,前列腺特异性膜抗原;,FCY1;,HSVTK;,前列腺肿瘤,0引言,1材料和方法,2结果,3讨论,【

     Construction of PSMAIRESFCY1TK eukaryotic expression vector and its expression in prostate cancer cell line

    YIN Ying1, LU Fan2, SU MingQuan1, HAO XiaoKe1

    1Department of Clinical Laboratories,Xijing Hospital, 2Department of Biochemistry & Molecular Biology, school of Basic Medicine.Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To construct the eukaryotic expression vector that specifically drives FCY1/TK double suicide gene by prostatespecific membrane antigen promoter(PSMAP)and to investigate the toxic effects of single or double gene on transfected prostate cancer cell line LNCaP after prodrug treatment.METHODS: Genomic DNA was extracted from benign prostate tissues and prostatespecific membrane antigen promoter(PSMAP) fragment was obtained by polymerase chain reaction (PCR).The pCMV promoter on eukaryotic expression vector pIRES was replaced by PSMA promoter and PSMAPIRES vector was obtained. The recombinant plasimd PSMAPIRES was identified by restriction analysis and DNA sequence. Then the gene FCY1 and TK was cloned into corresponding sites of PSMAIRES and the pPSMAIRESFCY1TK eukaryotic expression vector was obtained.Lipofection- mediated genetransfers were performed with the obtained vector and a control plasmid, pIRES, in human prostate cancer cell line LNCaP cells.RTPCR was used to demonstrate successful transcription.The toxic effects of 5FC, GCV, and both of them on transfected LNCaP cells were explored by MTT assay.RESULTS: A 1400 bp DNA fragment was amplified with PCR.Sequence and restriction analysis showed that PSMA promoter by amplified was identical with that of GenBank reported(GenBank accession number AF007544). The expression of suicide genes TK and FCY1 could be detected by RTPCR.The growth inhabition rate (GIR) in double gene group were higher than that of single gene.(P<005)CONCLUSION: pPSMAIRESFCY1TK double suicide gene system has significant toxic effect on LNCaP cells in vitro, which was superior. ......

您现在查看是摘要页,全文长 11020 字符
如果您在使用手机等流览时无法查看或下载全文,可能是被搜索引擎失真“转码”,请点击屏幕最下方的“电脑版”或“原网页”访问。


限于服务器压力,网站部分信息只供爱心会员或有一定积分的注册会员流览。
此信息需要 2 积分(免费注册登录后每天可以领取10个积分)。
用户名
密 码
  忘了密码

如果您还不是100md.com会员,欢迎 免费注册
如果您想获得积分,点击这里 查看积分规则
如果您想订购杂志,请直接与《第四军医大学学报》编辑部联系。
    信息仅供参考,不构成任何之建议、推荐或指引。文章版权属于原著作权人,若您认为此文不宜被收录供大家免费阅读,请邮件或电话通知我们,我们收到通知后,会立即将您的作品从本网站删除。

   微信文章  关注百拇  评论几句  搜索更多