Mavilio F, Sposi N M, Petrini M, Bottero L, Marinucci M, De Rossi G, Amadori S, Mandelli F, Peschle C
Proc Natl Acad Sci U S A. 1986 Jun;83(12):4394-8. doi: 10.1073/pnas.83.12.4394.
The structure and the expression of 11 cellular oncogenes (protooncogenes) were analyzed in primary cells from 20 acute lymphocytic (ALL) and 31 acute myelogenous (AML) leukemia patients. Neoplastic cells, obtained prior to initiation of therapy, were purified and classified, on the basis of both surface antigen pattern and morphology, into pre-B, B, and T ALL and M1-M5 AML. RNA was extracted and analyzed for expression of cellular oncogenes coding for nuclear proteins (c-myc, c-myb, c-fos), the beta-chain of platelet-derived growth factor (c-sis), growth factor receptors or related proteins (c-src, c-abl, c-fes, c-erbB), or putative intermediate transducers of mitogenic signals (c-Ha-ras, c-Ki-ras, c-N-ras). Quantitative analysis of total RNA was carried out by dot blot hybridization to specific cDNA or genomic probes. Number and size of transcripts were evaluated by blot hybridization of electrophoretically fractionated poly(A)+ RNA. Expression of c-myc and c-myb was detected in all leukemic cells at variable levels and was characterized by well-defined patterns within ALL subtypes. Conversely, significant levels of c-fos transcripts were detected only in myelomonocytic (M4) and monocytic (M5) leukemias. Among the "src-family," c-fes was expressed more in AML than ALL, and c-abl was expressed at variable but not elevated levels in all leukemia types. c-Ha-ras was uniformly expressed at low levels, as in non-neoplastic cells. c-Ki-ras transcription was detected only in T ALL; N-ras expression was barely demonstrable. The structure of these protooncogenes was not grossly modified, as evaluated by Southern analysis, except for c-myc rearrangement in B ALL. These studies indicate that cellular oncogene expression in specific subtypes of leukemic cells may relate to either the proliferative activity (c-myc, c-myb) or the differentiation state (c-fos) of the cells, or possibly to expression of receptors for putative hemopoiesis-related growth factors (c-fes, c-abl). Our data provide a basis for in-depth analysis of protooncogene expression in normal and neoplastic hemopoiesis.
对20例急性淋巴细胞白血病(ALL)和31例急性髓细胞白血病(AML)患者的原代细胞中11种细胞癌基因(原癌基因)的结构和表达进行了分析。在治疗开始前获取的肿瘤细胞,根据表面抗原模式和形态学进行纯化和分类,分为前B、B和T-ALL以及M1-M5 AML。提取RNA并分析编码核蛋白(c-myc、c-myb、c-fos)、血小板衍生生长因子β链(c-sis)、生长因子受体或相关蛋白(c-src、c-abl、c-fes、c-erbB)或有丝分裂信号假定中间转导子(c-Ha-ras、c-Ki-ras、c-N-ras)的细胞癌基因的表达。通过与特异性cDNA或基因组探针的点杂交对总RNA进行定量分析。通过对电泳分离的聚腺苷酸加尾RNA进行印迹杂交来评估转录本的数量和大小。在所有白血病细胞中均检测到c-myc和c-myb的表达,其水平各不相同,并且在ALL亚型中具有明确的模式。相反,仅在粒单核细胞白血病(M4)和单核细胞白血病(M5)中检测到显著水平的c-fos转录本。在“src家族”中,c-fes在AML中的表达高于ALL,而c-abl在所有白血病类型中的表达水平各不相同,但均未升高。c-Ha-ras在非肿瘤细胞中一样,均以低水平持续表达。仅在T-ALL中检测到c-Ki-ras转录;几乎未检测到N-ras表达。通过Southern分析评估,除了B-ALL中的c-myc重排外,这些原癌基因的结构没有明显改变。这些研究表明,白血病细胞特定亚型中的细胞癌基因表达可能与细胞的增殖活性(c-myc、c-myb)或分化状态(c-fos)有关,或者可能与假定的造血相关生长因子受体的表达(c-fes、c-abl)有关。我们的数据为深入分析正常和肿瘤造血中原癌基因的表达提供了基础。