目的：构建携蜂毒素基因及甲胎蛋白（AFP）启动子（rAFP）的重组腺病毒载体，探讨rAFP驱动的蜂毒素基因在体外对肝癌细胞的特异性杀伤作用。方法：人工合成蜂毒素基因，置于rAFP之后，用细菌内高效同源重组法将目的基因重组入腺病毒质粒中，转染人胚肾293细胞进行腺病毒包装；采用MTT法测定蜂毒素基因转染对AFP阳性、阴性肝癌细胞及正常肝细胞增殖的影响。结果：携蜂毒素基因重组腺病毒载体构建成功，MTT实验证明携蜂毒素基因重组腺病毒转染后，AFP阳性肝癌细胞的增殖受到明显抑制，而对AFP阴性肝癌细胞及正常肝细胞无明显影响；无蜂毒素基因的重组腺病毒转染，则对各种细胞增殖均无抑制作用。结论：表达rAFP驱动的蜂毒素基因的重组腺病毒在体外可特异性地杀伤AFP阳性肝癌细胞。

Objective:To evaluate the inhititory effects of the recombinant adenovirus vector expressing melittin gene under the control of human α-fetoprotein (AFP).Methods:The melittin gene was inserted into adenoviral vector through a bacterial homologous recombinant system, then the recombinant adenoviral vector was transfected to 293 cells to generate the desired recombinant adenoviruses. AFP-positive or AFP-negative hepatocarcinoma cells or hepatic cells were infected respectively with the recombinant adenoviruses and the proliferative inhibition was evaluated by MTT method.Results:Recombinant adenovirus containing melittin gene and AFP promoter was constructed successfully. When infected with the recombinant adenoviruses the inhibition of proliferative was observed in AFP-positive hepatocarcinoma cells, but not in AFP-negative hepatocarcinoma cells. Conclusions: Recombinant adenovirus expressing melittin gene under the control of human AFP promoter could specifically inhibit the proliferation of AFP-positive hepatocarcinoma cells.

R735.7

军队“九五”重点课题（98JD3269）；上海市科技发展基金（98XD14020）； 上海市“百人计划”（97BR044）