Ward M, Richardson C, Pioli P, Smith L, Podda S, Goff S, Hesdorffer C, Bank A
Department of Medicine, Columbia University New York, NY 10032.
Blood. 1994 Sep 1;84(5):1408-14.
The human multiple-drug resistance (MDR1) gene has been transferred into human hematopoietic progenitors using retroviral gene transfer. Human bone marrow cells and isolated CD34+ cells isolated from marrow were exposed to growth factors interleukin-3 (IL-3), IL-6, and stem cell factor for 48 hours and then to two changes of MDR retroviral supernatants over the next 24 hours. Progenitor assays in methylcellulose at this time showed that 18% to 70% of BFU-E and 30% to 60% of CFU-GM contain the transferred MDR gene by polymerase chain reaction analysis. Up to 11.2% of the progeny of these cells express increased amounts of MDR glycoprotein on their surface by fluorescence-activated cell sorter (FACS) analysis. In addition, transduced cells are enriched in high MDR-expressing cells after exposure to taxol as assessed by FACS analysis, and by resistance of BFU-E to taxol (Bristol-Myers Squibb, Princeton, NJ). These studies indicate the feasibility of using MDR gene transfer as a means of enriching marrow for MDR-transduced cells. They also provide the basis of a phase 1 clinical protocol in patients with advanced cancers not involving the bone marrow for the use of MDR gene transfer as a means of protecting marrow cells, which normally express low levels of MDR, from the myelosuppressive effects of drugs like taxol.
利用逆转录病毒基因转移技术，已将人类多药耐药（MDR1）基因转入人类造血祖细胞。将人类骨髓细胞以及从骨髓中分离出的CD34+细胞暴露于生长因子白细胞介素-3（IL-3）、IL-6和干细胞因子中48小时，然后在接下来的24小时内两次更换MDR逆转录病毒上清液。此时在甲基纤维素中进行的祖细胞分析表明，通过聚合酶链反应分析，18%至70%的爆式红系集落形成单位（BFU-E）和30%至60%的粒-巨噬细胞集落形成单位（CFU-GM）含有转移的MDR基因。通过荧光激活细胞分选仪（FACS）分析，这些细胞的后代中高达11.2%在其表面表达增加量的MDR糖蛋白。此外，通过FACS分析以及BFU-E对紫杉醇（百时美施贵宝公司，新泽西州普林斯顿）的抗性评估，转导细胞在暴露于紫杉醇后富含高表达MDR的细胞。这些研究表明，使用MDR基因转移作为富集骨髓中MDR转导细胞的一种手段是可行的。它们还为一项1期临床方案提供了基础，该方案针对不涉及骨髓的晚期癌症患者，使用MDR基因转移作为保护通常表达低水平MDR的骨髓细胞免受紫杉醇等药物骨髓抑制作用的一种手段。
