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Construction of a targeting adenoviral vector carrying AFP promoter for expressing EGFP gene in AFP-producing hepatocarcinoma cell 被引量:21
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作者 Yu-JunShi Jian-PingGong +4 位作者 Chang-AnLiu Xu-HongLi YingMei canmi Yan-YingHuo 《World Journal of Gastroenterology》 SCIE CAS CSCD 2004年第2期186-189,共4页
AIM:To construct a recombinant adenoviral vector carrying AFP promoter and EGFP gene for specific expression of EGFP gene in AFP producing hepatocellular carcinoma (HCC) HepG2 cells.METHODS: Based on the Adeno-X^TM ex... AIM:To construct a recombinant adenoviral vector carrying AFP promoter and EGFP gene for specific expression of EGFP gene in AFP producing hepatocellular carcinoma (HCC) HepG2 cells.METHODS: Based on the Adeno-X^TM expression system, the human immediate early cytomegalovirus promoter (PCMV IE)was removed from the plasmid, pshuttle,and replaced by a 0.3 kb (α-fetoprotein (AFP) promoter that was synthesized by polymerase chain reaction (PCR).The enhanced green fluorescent protein (EGFP) gene was inserted into the multiclone site (MCS),and then the recombinant adenovirus vector carrying the 0.3kb AFP promoter and EGFP gene was constructed.Cells of a normal liver cell line (LO2),a hepatocarcinoma cell line (HepG2) and a cervical cancer cell line (HeLa) were transfected with the adenovirus.Northern blot and fluorescence microscopy were used to detect the expression of the EGFP gene at mRNA or protein level in three different cell lines.RESULTS:The 0.3kb AFP promoter was synthesized through PCR from the human genome.The AFP promoter and EGFP gene were directly inserted into the plasmid pshuttle as confirmed by restriction digestion and DNA sequencing.Northern blot showed that EGFP gene was markedly transcribed in HepG2 cells,but only slightly in LO2 and HeLa cells.In addition,strong green fluorescence was observed in HepG2 cells under a fluorescence microscopy,but fluorescence was very weak in LO2 and HeLa cells.CONCLUSION:Under control of the 0.3kb human AFP promoter, the recombinant adenovirus vector carrying EGFP gene can be specially expressed in AFP-producing HepG2 cells,Therefore,this adenovirus system can be used as a novel,potent and specific tool for gene-targeting therapy for the AFP positive primary hepatocellular carcinoma, 展开更多
关键词 肝细胞癌 聚合酶链反应 AFP EGFP基因 基因序列 重组肾病毒受体
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