激光杂志
激光雜誌
격광잡지
LASER JOURNAL
2011年
5期
73-75
,共3页
多药耐药基因%转染%造血细胞
多藥耐藥基因%轉染%造血細胞
다약내약기인%전염%조혈세포
muhidmg resistance gene%taansfection%hematopoietic cells
目的：探讨mdrl基因体外转染兔骨髓造血细胞的条件及转染后在细胞中的功能性表达。方法：秋水仙碱（90ng／ml）筛选含人mdrlcDNA的产病毒包装细胞PA317-HaMDR1／A，制备浓缩病毒上清液并转染兔骨髓单个核细胞；分别用PCR法、免疫组织化学法和柔红霉素（daunorubicinDNR）排出试验检测mdr1基因的整合及功能性表达。结果：经秋水仙碱筛选后，产病毒包装糖蛋白（P—glycoprotein P-gp）表达增强；外源性mdrl基因能成功的导人兔骨髓造血细胞，转染2日、4日的转染率分别为22％、37％；转入的mdr1基因能发挥药物外排泵的功能。结论：采用浓缩病毒上清转染法能成功的将外源性mdr1基因导人兔骨髓造血细胞中并获得稳定的功能性表达，为进一步研究mdr1基因转染骨髓造血细胞后自体回输在大剂量化疗中对骨髓保护作用的研究提供了依据。
目的：探討mdrl基因體外轉染兔骨髓造血細胞的條件及轉染後在細胞中的功能性錶達。方法：鞦水仙堿（90ng／ml）篩選含人mdrlcDNA的產病毒包裝細胞PA317-HaMDR1／A，製備濃縮病毒上清液併轉染兔骨髓單箇覈細胞；分彆用PCR法、免疫組織化學法和柔紅黴素（daunorubicinDNR）排齣試驗檢測mdr1基因的整閤及功能性錶達。結果：經鞦水仙堿篩選後，產病毒包裝糖蛋白（P—glycoprotein P-gp）錶達增彊；外源性mdrl基因能成功的導人兔骨髓造血細胞，轉染2日、4日的轉染率分彆為22％、37％；轉入的mdr1基因能髮揮藥物外排泵的功能。結論：採用濃縮病毒上清轉染法能成功的將外源性mdr1基因導人兔骨髓造血細胞中併穫得穩定的功能性錶達，為進一步研究mdr1基因轉染骨髓造血細胞後自體迴輸在大劑量化療中對骨髓保護作用的研究提供瞭依據。
목적：탐토mdrl기인체외전염토골수조혈세포적조건급전염후재세포중적공능성표체。방법：추수선감（90ng／ml）사선함인mdrlcDNA적산병독포장세포PA317-HaMDR1／A，제비농축병독상청액병전염토골수단개핵세포；분별용PCR법、면역조직화학법화유홍매소（daunorubicinDNR）배출시험검측mdr1기인적정합급공능성표체。결과：경추수선감사선후，산병독포장당단백（P—glycoprotein P-gp）표체증강；외원성mdrl기인능성공적도인토골수조혈세포，전염2일、4일적전염솔분별위22％、37％；전입적mdr1기인능발휘약물외배빙적공능。결론：채용농축병독상청전염법능성공적장외원성mdr1기인도인토골수조혈세포중병획득은정적공능성표체，위진일보연구mdr1기인전염골수조혈세포후자체회수재대제양화료중대골수보호작용적연구제공료의거。
Objective： To explore the optimal transfection conditions in which the-foreign mdrl gene is transferred into hematopoietic cells of rabbit bone marrow, and test the expression and function of the mdrl gene in the ceils. Methods： An amphotropic virus producer cell line, PA317 - HaMDR l/A, which contains a full- length cDNA of human mdrl gene, was screened with colehicines（90ng/ml）,and the supernatant was collected and concentrated to cocultivated with the bone marrow mononuclear cells of the rabbit. The integratian, transfection rate and physiological function of mdr 1 gene was tested by PCR, SP immunohistochemical method and DNR extrusion test respectively. Results：After screening by colchicinesj the expression of P- gly, coprotein （p- gp）of PA317 - HaMDR1/A was enhanced. The foreign mdr 1 gene was transferred into hematopoietic ceils of rabbit bone marrow successfully, and the transduction rate of 2 days and 4 days were 22 %, 37 %. The transferred mdr 1 gene had it＇ s physiological function. Conclusion ： The foreign mdr 1 gene can be transferred into hematopoietic cells of rabbit bone marrow successfully by concentrated virus supematant transfection method, and the transferred gene can be expressed stably and effectively. The study has provided a basis for the further research on chemopmtection experiment of the mdr 1 gene transferred into the bone marrow cells.
