IL-3-PE38KDEL转染CIK细胞对HL60细胞体内外杀伤作用的实验研究

摘要

目的 研究IL-3-PE38KDEL基因转染细胞因子诱导的杀伤细胞(CIK)对HL60白血病细胞的杀伤作用.方法 应用脂质体将融合基因IL-3-PE38KDEL转染CIK细胞,酶联免疫吸附试验(ELISA)和流式细胞术(FCM)法检测转基因前后CIK分泌细胞因子能力及细胞表型的变化,MTS法检测基因转染CIK细胞对HL60细胞细胞毒活性的变化,建立HL60裸鼠模型,给予IL-3-PE38KDEL转染CIK细胞、空质粒转染CIK细胞、未转染CIK细胞及生理盐水,观察其对裸鼠移植瘤生长的抑制作用.结果 通过脂质体成功将IL-3-PE38KDEL转染CIK细胞,转染前后CIK细胞的分泌细胞因子能力及细胞表型无明显变化,转染后CIK细胞对HL60细胞的杀伤活性较空质粒转染CIK细胞及未转染CIK细胞明显提高,体内实验表明基因转染CIK细胞可明显抑制裸鼠皮下HL60细胞移植瘤的生长.结论 IL-3-PE38KDEL基因转染CIK细胞能够明显抑制体内外HL60细胞的生长,同时不影响CIK分泌细胞因子能力及细胞表型.%Objective To investigate the anti-tumor effect of cytokine induced-killer (CIK) cells transfected with IL-3-PE38KDEL fusion gene on HL60 cells in vitro and in vivo.Methods IL-3-PE38KDEL fusion gene was transfected into CIK cells by liposome mediate.The phenotype of CIK cells was detected by flow cytometry.The concentration of IFN-γ and TNF-α in supernatant of CIK cells was determined by enzyme-linked immunosorbent assay (ELISA).Cytotoxicity of CIK cells transfected with IL3-PE38KDEL gene against HL60 cells was detected by MTS assay.The human leukemia HL60 cell line subcutaneous xenograft tumor model in nude mice was established.CIK cells transfected with IL-3-PE38KDEL,CIK cells transfected with empty vector,non-transfeced CIK cells and normal saline were given respectively to study the growth inhibitory effect on the transplanted tumor in nude mice.Results The phenotype and the level of cytokine remained unaltered in the IL-3-PE38KDEL gene transfected CIK cells.By comparison with non-transfected CIK cells and those transfected by empty vector,CIK cells transfected with IL-3-PE38KDEL had a significantly high anti-tumor cytotoxic activity against HL60 cells in vitro,and decreased tumor weight.Conclusion IL-3-PE38KDEL fusion gene transfection can enhance the anti-tumor activity of CIK cells.