Engelhard H H, Butler A B, Bauer K D
Department of Pathology, Northwestern Memorial Hospital, Northwestern University, Chicago, Illinois.
J Neurosurg. 1989 Aug;71(2):224-32. doi: 10.3171/jns.1989.71.2.0224.
The identification of a quantifiable oncoprotein marker in glial cells could lead to its use as an aid in the diagnosis, grading, and treatment of tumours of glial origin. In this study, monoclonal antibodies to the c-myc oncoprotein were used in conjunction with immunofluorescence microscopy, flow cytometry, and immunoblot analysis to quantitate and characterize the expression of this oncoprotein in neoplastic and benign cultured glial cells and brain-tumor tissue. Flow cytometric analysis revealed that the c-myc oncoprotein was highly expressed in neoplastic cell lines and in glioblastoma tumor specimens. In contrast, anti-c-myc oncoprotein staining was not present in a non-neoplastic glial cell line or in a benign brain tissue specimen. Immunoblot analysis revealed two distinct c-myc oncoprotein bands, having molecular weights of 64 and 75 kD. Densitometric determinations of the relative levels of the 64-kD protein were in good agreement with the determinations made by flow cytometry. Flow cytometry was also used to relate the quantity of the c-myc oncoprotein present in the cells to their cell cycle phase. In the malignant cultured cells, the protein underwent an approximate twofold increase as the cells progressed from G1/G0 to G2/M in the cell cycle. The present results suggest that the c-myc oncoprotein may prove to be a useful marker for the proliferation status and/or malignancy of glial cells.
在神经胶质细胞中鉴定出一种可量化的癌蛋白标志物,可能会使其有助于神经胶质源性肿瘤的诊断、分级和治疗。在本研究中,针对c-myc癌蛋白的单克隆抗体与免疫荧光显微镜、流式细胞术和免疫印迹分析结合使用,以定量和表征该癌蛋白在肿瘤性和良性培养的神经胶质细胞以及脑肿瘤组织中的表达。流式细胞术分析显示,c-myc癌蛋白在肿瘤细胞系和胶质母细胞瘤肿瘤标本中高表达。相比之下,抗c-myc癌蛋白染色在非肿瘤性神经胶质细胞系或良性脑组织标本中不存在。免疫印迹分析显示出两条不同的c-myc癌蛋白条带,分子量分别为64 kD和75 kD。对64-kD蛋白相对水平的光密度测定与流式细胞术测定结果高度一致。流式细胞术还用于将细胞中存在的c-myc癌蛋白量与其细胞周期阶段相关联。在恶性培养细胞中,随着细胞在细胞周期中从G1/G0期进展到G2/M期,该蛋白含量大约增加了两倍。目前的结果表明,c-myc癌蛋白可能被证明是神经胶质细胞增殖状态和/或恶性程度的有用标志物。