白细胞介素2基因转移的肿瘤细胞克隆的建立及其体内外生长特性的研究

IN VITRO AND IN VIVO GROWTHCHARACTRERISTICS OF ACLONE OF TUMOR CELLS TRANSFECTED WITH IL-2 GENE

为了开展ＩＬ－２基因转移后免疫原性增强的新型瘤苗抗肿瘤作用的研究，须首先获得ＩＬ－２基因转移后、能够高分泌ＩＬ－２的肿瘤细胞并确定其生长特性。本室构建了含５６３ｂｐ的小鼠ＩＬ－２全长ｃＤＮＡ的真核表达载体ＢＭＧＮｅｏ－ＩＬ－２，采用磷酸钙ＤＮＡ共沉淀法将ＢＭＧＮｅｏ－ＩＬ－２转移入Ｂ１６黑色素瘤细胞中，通过Ｇ４１８抗性筛选、有限稀释法及活性检测，获得一株高分泌ＩＬ－２（５０６Ｕ／ｍｌ）克隆，并经Ｓｏｕｔｈｅｒｎ印迹法证实ＩＬ－２基因已转入该克隆中。结果表明ＩＬ－２基因转移的肿瘤细胞体外生长能力没有明显变化，但体内致瘤原性显著下降，从而为制备新型瘤苗打下了基础。

n order to investigate the antitumor effect of tumor vaccine transfected with IL- 2 genewhich has increased immunogenicity,IL- 2 gene- transfected tumor cells which secrete high Ievelof IL-2 must be obtained and the growth characteristics of these tumor cells should be identified. The expression vector BMGNeo-IL-2 carrying 563 bp full-Iength mlL-2 cDNA was constructedand was transfected into B16 melanoma cells by calcium phosphate coprecipitation method. AfterG418 resistance selection,limiting dilution and IL-2 assay, a clone secreting highest Ievel of IL-2(506 U/ml)was selected,and then identified by Southern blot. Our results demonstrated thatIL-2 gene-transfected tumor cells exhibited unchanged in vitro growth but decreased tumori-genicity.A new type of tumor vaccine could be prepared form the IL-2-secreting tumor cell cloneestablished in the experiment.