Construction of a targeting adenoviral vector carrying AFP promoter for expressing EGFP gene in AFP-producing hepatocarcinoma cell

作     者：Yu-JunShi Jian-PingGong Chang-AnLiu Xu-HongLi YingMei CanMi Yan-YingHuo 

作者机构：DepartmentofGeneralSurgerytheSecondCollegeofClinicalMedicineandtheSecondAffiliatedHospitalofChongqingUniversityofMedicalSciencesChongqing400010China DepartmentofPathoiogyBasicMedicalCollegeChongqingUniversityofMedicalSciencesChongqing400016China InstituteforRadiologyMilitaryAcademyofMedicalSciencesBeijing100850China 

出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)

年 卷 期：2004年第10卷第2期

页      面：186-189页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：Supported by the Key Program of Medical Science Foundation of Chongqing Public Health Bureau  01-1-018 

主　　题：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 

摘      要：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 ***: 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 *** of a normal liver cell line (LO2),a hepatocarcinoma cell line (HepG2) and a cervical cancer cell line (HeLa) were transfected with the *** blot and fluorescence microscopy were used to detect the expression of the EGFP gene at mRNA or protein level in three different cell ***:The 0.3kb AFP promoter was synthesized through PCR from the human *** AFP promoter and EGFP gene were directly inserted into the plasmid pshuttle as confirmed by restriction digestion and DNA *** blot showed that EGFP gene was markedly transcribed in HepG2 cells,but only slightly in LO2 and HeLa *** addition,strong green fluorescence was observed in HepG2 cells under a fluorescence microscopy,but fluorescence was very weak in LO2 and HeLa ***: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,