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Transformation of type 1 astrocytes with N‐ethyl‐N‐nitrosourea: Establishment of an in vitro system and the role of the p53 gene
Author(s) -
Hiraga Shoju,
Arita Norio,
Ohnishi Takanori,
Izumoto Shuichi,
Taki Takuyu,
Yamamoto Hiroshi,
Higuchi Masahide,
Hayakawa Toru
Publication year - 1995
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/glia.440130107
Subject(s) - biology , carcinogenesis , astrocyte , in vitro , transformation (genetics) , microbiology and biotechnology , neuroglia , mutant , immunohistochemistry , western blot , cell culture , nitrosourea , gene , genetics , immunology , central nervous system , endocrinology , chemotherapy
N‐ethyl‐N‐nitrosourea (ENU)‐induced gliomas, animal models of human gliomas, are most frequently oligodendrocytic, while human gliomas tend to be astrocytic. To facilitate a detailed study of human glial carcinogenesis, we developed an in vitro system using type 1 astrocyte transformation with ENU. Type 1 astrocytes from fetal Wistar rat brain were treated by a single dose of ENU. Transformed colonies appeared 50 days after exposure to single doses of ENU greater than 150 μg/mL. Cloned cells from these colonies retained the immunohistochemical characteristics of type 1 astrocytes. They showed rapid growth and high saturation densities, colony formation in low (2%) serum medium and gave rise to tumors when injected into nude mice. When p 53 expression was studied at each passage, a single cell positive for mutant p 53 protein emerged 40 days after ENU treatment. In the next 1–3 passages, the mutant p 53 positive cell formed piled‐up colonies and exhibited dominant growth. Northern blot analysis showed markedly increased accumulations of p 53 mRNA in transformed cells. This in vitro transformation system of type 1 astrocytes provides a valuable tool for further investigations of astrocyte carcinogenesis. © 1995 Wiley‐Liss, Inc.