细胞因子对小鼠造血祖细胞基因转移效率的影响  

Efects of cytokines on efficiency of gene transfer into murine hematopoletic progenitors

作　　者：宋献民[1] 杨建民[1] 闵碧荷[1] 卢大儒[1] 李川晟[1] 孟沛霖[1] 

机构地区：[1]第二军医大学长海医院血液内科,复旦大学遗传学研究所

出　　处：《第二军医大学学报》1996年第1期10-14,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金

摘　　要：目的：观察细胞因子对逆转录病毒载体介导的ＮｅｏＲ基因在小鼠造血祖细胞及人Ｋ５６２细胞中基因转移效率的影响，以探求最有效地促进造血细胞基因转移的细胞因子或其组合。方法：白细胞介素１α（ＩＬ－１α）、白细胞介素３（ＩＬ－３）、干细胞因子（ＳＣＦ）作用下预培养的骨髓细胞用含病毒颗粒的上清转染后，经Ｇ４１８或不含Ｇ４１８体外半固体培养，以生物学方法及ＰＣＲ方法检测造血祖细胞的某因转移效率。结果：ＩＬ－１α／ＩＬ－３／ＳＣＦ联合应用能最有效地提高小鼠ＣＦＵ－ＧＭ基因转移效率，使转移效率从（６．０４±１．３４）％提高到（４３．６±５．９４）％；ＳＣＦ单独应用能最有效地促进Ｋ５６２细胞基因转移，使其转移效率从（１９．０４±１．５８）％提高到（５４．４６±２．１３）％。结论：ＩＬ－１α／ＩＬ－３／ＳＣＦ联合应用可提高造血干、祖细胞基因转移效率，ＳＣＦ可用于急性髓性白血病患者自体骨髓移植时基因标志的临床研究。Objective:To observe the effects of cytokines on the efficiency of gene transfer into murine hematopoietic progenitors and human K562 cells mediated by retrovirus (RV) containing bacterial neomycin resistant gene (NeoR). Methods: Cytokines were used in bone marrow cells preincubation and retrovirus vectors-containing supernant transfection. The transfected cells were planted in semisoloid culture with or without G418. Gene transfer efficiency into hematopoietic progenitors was evaluated by biological assay and PCR analysis. Results:The most efficient combination of cytokines,IL-Iα/IL-3/SCF,increased the efficiency of gene transfer into murine CFU-GM from (6.04±1.34)% to(43.6±5.94)%.SCF alone most efficiently facilitated the gene transfer into human K562 cells and increased the gene transfer efficiency from (19. 04±1.58)% to(54. 46±2.13)%.Conclusion: The combination of IL-1α/IL-3/SCF can increase the efficiency of gene transfer into hematopoietic stem cells (HSC) and progenitors,and in the treatment of acute myeloid leukemia (AML) by autologous bone marrow transplantation(ABMT),SCF can facilitate gene transfer into hematopoietic cells in gene- marking clinical studies.

关 键 词：细胞因子 基因转移效率 造血干细胞 

分 类 号：Q75[生物学—分子生物学]