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人胶质母细胞瘤细胞体外VI型胶原蛋白表达的免疫荧光和生化研究。

Immunofluorescence and biochemical studies of the type VI collagen expression by human glioblastoma cells in vitro.

作者信息

Han J, Daniel J C, Lieska N, Pappas G D

机构信息

Department of Anatomy & Cell Biology, College of Medicine, University of Illinois, Chicago 60612.

出版信息

Neurol Res. 1994 Oct;16(5):370-5. doi: 10.1080/01616412.1994.11740256.

Abstract

The human glioblastoma cell line U-87 MG was found to express a 140 kD polypeptide which was recognized on immunoblot analysis by a monoclonal antibody to type VI collagen. This polypeptide was digestible by a highly purified bacterial collagenase. After treatment of U-87 MG cells by pepsin, the protein profile revealed the two major pepsin-resistant fragments identical in Mr to those of collagen VI extracted from human placenta. The respective peptide maps from V8 protease one-dimensional gels of these two fragments were identical to those obtained with human collagen VI. Immunofluorescent staining by antibodies to type VI collagen was observed in the extracellular matrix. Moreover, U-87 MG cells were found to be positive for A2B5, a cell surface marker specific for O-2A type glial precursor cells. These data indicate that the human glioblastoma cell line U-87 MG exhibits the properties of glial precursor cells and expresses collagen type VI in vitro. This cell line therefore may prove valuable for comparative investigations of the regulation of type VI collagen synthesis, and may be useful as a model to study the function and pathological importance of type VI collagen in human brain tumours, both in vitro and in vivo.

摘要

人类胶质母细胞瘤细胞系U - 87 MG被发现表达一种140 kD的多肽,在免疫印迹分析中,该多肽可被抗VI型胶原的单克隆抗体识别。这种多肽可被高度纯化的细菌胶原酶消化。用胃蛋白酶处理U - 87 MG细胞后,蛋白质谱显示出两个主要的抗胃蛋白酶片段,其分子量与从人胎盘中提取的VI型胶原的片段相同。这两个片段在V8蛋白酶一维凝胶上各自的肽图与用人VI型胶原获得的肽图相同。在细胞外基质中观察到抗VI型胶原抗体的免疫荧光染色。此外,发现U - 87 MG细胞对A2B5呈阳性,A2B5是O - 2A型胶质前体细胞特有的细胞表面标志物。这些数据表明,人类胶质母细胞瘤细胞系U - 87 MG在体外表现出胶质前体细胞的特性并表达VI型胶原。因此,该细胞系可能对VI型胶原合成调控的比较研究有价值,并且可能作为一个模型,用于在体外和体内研究VI型胶原在人脑肿瘤中的功能和病理重要性。

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