外源性IL-3cDNA导入骨髓基质细胞及对造血的调控作用  

Bone marrow stromal cells transferred by exogenous IL-3 cDNA and its regulatory ef- fect on hematopoiesis in mice

作　　者：袁良平[1] 粟永萍[1] 程天民[1] 刘林[1] 郑华金[2] 王延江[1] 蒲晓允[3] 艾国平[1] 刘晓红 瞿红云 李青[2] 

机构地区：[1]第三军医大学预防医学系防原医学教研室,全军复合伤研究所 [2]第三军医大学成都军区成都总医院血液内科,成都610083 [3]第三军医大学医学检验系临床检验学教研室,重庆400038

出　　处：《第三军医大学学报》2001年第8期948-950,共3页Journal of Third Military Medical University

摘　　要：目的 探索将外源性ＩＬ－３ｃＤＮＡ导入骨髓基质细胞的可能性。方法 将小鼠ＩＬ－３ｃＤＮＡ克隆到逆转录病毒载体（ｐＬＸ－ＳＮ）上，然后将重组的ＩＬ－３ ｃＤＮＡ用脂质体转染到包装细胞ＰＡ３１７上，Ｇ４１８筛选后得到抗性克隆，将含病毒的上清液转染骨髓基质细胞。用自行设计的引物，检测ｐＬＸＳＮ上的ＮｅｏＲ基因及转入的ＩＬ－３基因。同时观察了转染ｐＬＸＳＮ／ＩＬ－３的基质细胞培养上清液对小鼠骨髓 ＣＦＵ－ＧＭ形成的影响，并测定其对造血因子依赖株 ＮＦＳ－６０细胞增殖活性的影响。结果 将含重组子 ｐＬＸＳＮ－ＩＬ－３和空载体ｐＬＸＳＮ的病毒培养上清感染 ＢＭＳ，前者用 ＰＣＲ扩增出５００ ｂｐ和１７０ ｂｐ（ＮｅｏＲ）的条带，后者仅扩增出 １７０ ｂｐ。转染ｐＬＸＳＮ／ＩＬ－３的基质细胞上清液有促进小鼠骨髓ＣＦＵ－ＧＭ增殖的作用，与空载体组及未转染组比较，有非常显著性差异（Ｐ＜０．０１）。结论 外源性的ＩＬ－３基因及ｐＬＸＳＮ上的ＮｅｏＲ已整合到骨髓基质细胞基因组ＤＮＡ中，并促进造血细胞增殖。Objective To investigate the feasibility of transferring exogenous IL-3 cDNA into bone marrow stromal cells (BMS). Methods Mouse IL-3 cDNA was cloned to retrovirus vector pLXSN and then the recombinant IL-3 cDNA was transfected on PA317 cells with lipofectin. The cells was selected with C418 after BMS was transfected by the supernatant containing virus. NeoR gene of pLXSN and transferred IL-3 gene in BMS were identified with PCR. CFU-GM of murine BMS culture medium and proliferative activity of hematopoietic factor-dependent cell line NFS-60 were observed in the supernatant of BMS with pLXSN/IL-3. Results IL-3 gene and NeoR gene of pLXSN were demonstrated in BMS transfected by the supernatant that containing pLXSN/IL-3, but only NeoR gene was seen in BMS containing pLXSn. The supernatant of BMS with pLXSN/IL-3 showed significant effects on the proliferation of CFU-GM in murine bone marrow, compared wiht that of the group BMS with pLXSN and the group BMS with- out pLXSN/IL-3 transfection. Conclusion The exogenous IL-3 and NeoR gene of pLXSN can be integrated into DNA genome of BMS, and promote the proliferation of hematopoietic cells.

关 键 词：白细胞介素-3 基质细胞 聚合酶链反应 造血细胞增殖 骨髓 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R331.22[医药卫生—基础医学]