pEGFP—AFP—TK真核表达载体的构建及其抗肝癌细胞效应

目的 构建AFP启动子介导的HSV／TK重组真核表达载体，并研究其荷瘤效应。方法PCR扩增AFP基因启动子、HSV—TK基因，将上述片断插入EGFP—N1载体的多克隆位点，从而构建甲胎蛋白启动子调控下HSV—TK基因重组表达载体pEGFP—AFP—TK。将其注射移植性H22肝癌模型C57BL／6J小鼠，并设转染空质粒pcDNA3．1组，观察两组的抗肿瘤效应。结果肝癌特异性真核表达载体pEGFP—AFP—TK构建成功，该真核表达载体免疫荷瘤小鼠后，pEGFP—AFP—TK组肿瘤细胞生长受到抑制，且该组生存期长于pcDNA3．1组（P〈0．05）。结论在受试动物体内可有效的诱导特异性抗肿瘤免疫应答。

Construction of pEGFP - AFP - TK eukaryotic recombinant plasmid and its effects of anti - hepatoma cells immunization

Objective To construct a recombinant eukaryotic expression vector containing （x - fetoprotein （AFP） promoter- mediated HSV/TK suicide gene, and evaluate its anti -tumor immunization effects in the mice bearing tumor. Methods The promoter of AFP gene, HSV - TK gene were amplified by RT- PCR. The gene segments mentioned above were cloned into the multiple cloning sites of the eukaryotic expression vector pEGFP -N1, pEGFP -AFP -TK, was constructed. The C57BL/6J mice model with transplantable H22 liver cancer, were randomly allocated into two groups as pcDNA3.1 and pEGFP- AFP- TK. The anti -tumor effects were observed. Results Hepatocellular carcinoma -specific eukaryotic expression vector carrying AFP promoter and HSV/TK suicide gene, pEGFP -AFP - TK, was sucessfully constructed. The growth of tumor was slower and the survival time was obviously longer of pEGFP - AFP - TK group than pcDNA3.1 group（ P 〈 0.05）. Conclusions The recombinant plasmid pEGFP- AFP- TK can induce anti -tumor immunization effects in the mice bearing tumor.