以逆转录病毒为载体的IL-2基因转染K562细胞的研究  

RETROVIRAL TRANSFER OF IL-2 GENE INTO HUMAN LEUKEMIA CELL LINE K562

作　　者：陈学良[1] 于燕霞[2] 刘春生[1] 张东青[1] 楚中华[1] 

机构地区：[1]山东大学齐鲁医院血液科 [2]山东大学第二医院血液科

出　　处：《山东医科大学学报》2001年第6期500-502,共3页Acta Academiae Medicinae Shandong

基　　金：山东省科委科研基金资助课题 ( 96 10 10 )

摘　　要：目的 :研究以逆转录病毒为载体将IL 2基因转入K5 62细胞及表达情况。方法 :应用脂质体介导的基因转移法 ,将质粒PGEN/IL 2导入包装细胞PA317,将K5 62细胞与PA317/IL 2细胞共培养 ,检测IL 2cDNA整合 ,mRNA转录情况 ,以IL 2依赖性CTLL 2细胞测定培养细胞上清中IL 2产量 ,比较转染前后细胞的体外生长情况和致瘤性。结果 :IL 2cDNA成功整合入K5 62细胞基因组中并稳定表达 ,K5 62 /IL 2细胞培养 2 4h上清中IL 2活性为 10 0U/ml,体内外观察均见细胞生长减慢。结论 :以脂质体介导的以双顺反子逆转录病毒为载体的基因转移方法 ,可成功地将人IL 2基因转入人白血病细胞株K5 62 ,并持续分泌高活性的IL 2 ,为下一步将IL 2基因转入CML造血细胞的研究打下基础。Objective:To transduce pGEN/IL 2 into K562 cells and investigate it's expression.Methods:Packaging cell PA317 were transfected with pGEN/IL 2 by liposome mediated gene transfer method.K562 cells were coincubated with PA317/IL 2 cells.Integration of IL 2 in the genome,transcription of its mRNA and expression of its protein in the transfected K562 cells were assayed by Southern blot,RT PCR and IL 2 dependent cell line CTLL.Results:IL 2 cDNA was integrated into K562 genome sucessfully.IL 2 mRNA was transcripted and IL 2 protein was expressed of 100u/10 6 cells per day by CTLL cell assay in the supernatant,K562 cells grew slower in vivo and in vitro.Conclusion:Lipofectin mediated bicistronic retrovirus gene transduction method is efficient.We Successfully transduced human IL 2 gene into packaging PA317 and human leukemia cell line K562 with this method and expressed high level of IL 2.

关 键 词：白细胞介素Ⅱ 基因转移 K562细胞 逆转录病毒 骨髓移植 

分 类 号：R457.7[医药卫生—治疗学] Q782[医药卫生—临床医学]