Keith W N, Brown R, Pragnell I B
CRC Department of Medical Oncology, Beatson Institute for Cancer Research, Glasgow, UK.
Br J Cancer. 1990 Sep;62(3):388-94. doi: 10.1038/bjc.1990.304.
Two retrovirus vectors were compared for their ability to express granulocyte-macrophage colony stimulating factor (GM-CSF) in a haematopoietic cell line, FDCP1, which is dependent on GM-CSF for survival. Both a MoMLV-based vector pVneoGM, and a MPSV-based vector, M3neoGM, were found to be capable of transmitting and expressing both GM-CSF and neomycin sequences in the myeloid FDCP1 cell line. Our results also demonstrate that pVneoGM is more efficient at generating GM-CSF independent colonies than M3neoGM. Analysis of cell lines derived after infection confirmed pVneoGM expressed higher levels of GM-CSF. Cell lines generated by infection with pVneoGM responded to levels of exogenous recombinant GM-CSF which did not stimulate growth of the parental cell line, suggesting autocrine stimulation may convey a proliferative advantage under sub-optimal growth conditions. Finally the parental vectors pVneo and M3neo were shown to be capable of expressing the neomycin gene in both murine haematopoietic progenitor and stem cells.
比较了两种逆转录病毒载体在造血细胞系FDCP1中表达粒细胞-巨噬细胞集落刺激因子(GM-CSF)的能力,该细胞系依赖GM-CSF存活。发现基于莫洛尼鼠白血病病毒(MoMLV)的载体pVneoGM和基于小鼠干细胞病毒(MPSV)的载体M3neoGM都能够在髓系FDCP1细胞系中传递和表达GM-CSF和新霉素序列。我们的结果还表明,pVneoGM在产生不依赖GM-CSF的集落方面比M3neoGM更有效。对感染后获得的细胞系的分析证实,pVneoGM表达的GM-CSF水平更高。用pVneoGM感染产生的细胞系对外源重组GM-CSF的水平有反应,而这些水平并未刺激亲代细胞系的生长,这表明自分泌刺激可能在次优生长条件下赋予增殖优势。最后,亲本载体pVneo和M3neo被证明能够在小鼠造血祖细胞和干细胞中表达新霉素基因。