ULBP3基因转染提高人外周血NK细胞对人成骨肉瘤Saos-2细胞的杀伤活性

ULBP3 gene transfection make Saos-2 cell line more sensitive to human NK killing

目的 观察ULBP3基因转染对人外周血NK细胞对Saos -2细胞杀伤活性的影响。方法 用RT -PCR方法从人成骨肉瘤组织中提取ULBP3基因 ,构建pcDNA3真核表达质粒 ,以Lipofectamine为载体转染人成骨肉瘤Saos-2细胞 ,用percoll密度梯度离心法分离人外周血NK细胞作为效应细胞 ,以Saos -2细胞和转染ULBP3基因的Saos-2细胞作为靶细胞 ,进行体外杀伤实验 ,比较两组的特异性杀伤率。结果 在效靶比为 3 0∶1时人外周血NK细胞对转染ULBP3基因的Saos-2细胞的特异性杀伤率明显高于对照组。结论 ULBP3基因转染能提高人外周血NK细胞对人成骨肉瘤Saos-2细胞的杀伤活性。

Objective To study the effect of ULBP3 gene transfection on the sensitivity of Saos-2 cell line to human NK killing.Methods The ULBP3 gene fragment obtained by RT-PCR from human osteosarcoma specimen was sequenced and cloned into the pCDNA3 eukaryotic expression vector.Then the recombinant plasmid was transfected into Saos-2 cell line with lipofectamine.With NK cell isolated from human peripheral blood as effect cell,the ULBP3 gene transfected Saos-2 cell and nontransfected Saos-2 cell labelled with 51Cr as target cell,the in vitro NK killing experiment was performed,and the rate of specific lysis was tested.Results The rate of specific lysis of ULBP3 gene transfected Saos-2 cell was much higher than that of nontransfected Saos-2 cell.Conclusion ULBP3 gene transfection make Saos-2 cell line more sensitive to human NK killing.