甲胎蛋白启动子驱动的蜂毒基因对肝癌细胞的特异性杀伤作用

The Inhibitive Effect of Melittin Gene under the Control of Human α-Fetoprotein Gene Promoter for Hepatocarcinoma Cells

目的 :构建携蜂毒素基因及甲胎蛋白 (AFP)启动子的重组腺病毒载体 ,探讨AFP启动子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 :采用细菌内高效同源重组法构建携蜂毒素基因及甲胎蛋白 (AFP)启动子的重组腺病毒载体 ,X gal染色测定重组腺病毒对肝癌细胞的转染效率 ,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 :重组腺病毒对肝癌细胞具有较高的转染率 ,MTT法证明携AFP启动子和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 ;含巨细胞病毒启动子和蜂毒素基因的重组腺病毒转染 ,则对AFP阳性及阴性肝癌细胞增殖均有抑制作用。结论

Objective:Construct recombinant adenovirus vector carrying melittin gene under the control of human α fetoprotein(AFP)gene promoter and study the special inhibition of the melittin gene for hepatocarcinoma cells in vitro.Methods:The recombinant adenovirus vectors carrying melittin gene under the control of AFP gene promoter were constructed through a new simplified bacterial homologous recombinant system,then the gene transduction efficiency for hepatocarcinoma cells was determined by X gal stain.After AFP positive or AFP negative hepatocarcinoma cells or hepatic cells were infected respectively with recombinant adenoviruses,the proliferative inhibition was evaluated by MTT method.Results:The recombinant adenovirus have higher gene transduction efficiency for hepatocacinoma cells.The proliferative inhibition was observed in AFP positive hepatocarcinoma cells infected with recombinant adenoviruses carrying melittin and AFP gene,but not in other cells.The proliferative inhibition was observed in AFP positive and AFP negitive hepatocarcinoma cells infected with recombinant adenoviruses carrying melittin and CMV gene.Conclusion:Melittin gene under the control of human AFP gene promoter can inhibit specially the proliferation of AFP positive hepatocarcinomacells.