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本文作者选用依赖于白细胞介素-3(IL-3)的造血细胞系32Dc13和NIH/3T3胚胎成纤维细胞作克隆亚系,利用霉酚酸耐受基因与癌基因连接转换技术,分离出高水平表达mRNA的亚克隆系,证明一些癌基因表达提高了辐射耐受性.这有助于解释血液肿瘤的复发机理.32Dc13细胞系来自C3H/HeJ小鼠的长期骨髓培养物,在IL-3克隆化体外传代培育数年,能形成中性粒细胞和肥大细胞集落.该细胞系对血红蛋白合成或红细胞生成素无反应.对IL-1、-2、-4、-5、-6、-7及G-CSF和GM-CSF并不发生增殖反应.用v-erb-B、v-sis、v-abl、v-src癌基因转染,以霉酚酸培养后,从该细胞中提取poly(A)~+mRNA,测试每个癌基因的表达水平.
The authors used the hematopoietic cell line 32Dc13 and NIH/3T3 embryonic fibroblasts, which are dependent on interleukin-3 (IL-3), as the clonal sublines, and used the mycophenolic acid tolerant gene and oncogene linkage conversion technology to isolate Levels of mRNA expression in subcloning lines demonstrate that some oncogene expressions increase radiation tolerance. This helps explain the mechanism of recurrence of hematologic tumors. 32Dc13 cell line is derived from long-term bone marrow culture of C3H/HeJ mice in IL-3 The cloned cells are passaged in vitro for several years to form neutrophils and mast cell colonies. This cell line is not responsive to hemoglobin synthesis or erythropoietin. For IL-1, -2, -4, -5, -6, - 7 and G-CSF and GM-CSF do not proliferate. Transfection with v-erb-B, v-sis, v-abl, v-src oncogene, cultured with mycophenolic acid, extraction from the cells Poly(A)~+mRNA was tested for the expression level of each oncogene.