Establishment of spontaneously immortalized rat type 1 astroglial cell lines: the role of p53 in astroglial carcinogenesis.
We established five spontaneously immortalized cell lines using purified rat type 1 astroglia on a rigid transfer schedule. All the cell lines maintained their polygonal shape, regular pavement growth, low saturation density, positive glial fibrillary acidic protein expression, and serum requirements, while none were tumorigenic in nude mice. We then obtained a spontaneously transformed cell line by maintaining the cells for 6 months at a high cell density. Since alterations of the tumor suppressor p53 gene have been reported in the immortalization of some cell lines and in transformation of others, we characterized p53 in immortalized, spontaneously transformed, and 5 Nethyl-N-nitrosourea (ENU)-transformed cell lines. While each of the ENU-induced or the spontaneously transformed cell lines exhibited p53 gene mutations that resulted in amino acid alterations, no alterations in the p53 gene were observed in any of the immortalized cell lines. Thus, alterations of the p53 protein correlate more strongly with transformation than with immortalization of type 1 astroglia. Immortalization may be regulated by gene(s) other than p53. Spontaneously immortalized type 1 astroglial cell lines may provide a new tool to investigate an initial step of astroglial carcinogenesis.
Department of Neurosurgery, Osaka University Medical School, Japan.
S
S
Hiraga
N
N
Arita
T
T
Ohnishi
S
S
Izumoto
T
T
Taki
M
M
Higuchi
K
K
Iwaisako
S
S
Sakoda
Y
Y
Yamamoto
T
T
Hayakawa
D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D001254:Astrocytoma / Q000503:physiopathology*; D015139:Blotting, Southern; D001932:Brain Neoplasms / Q000503:physiopathology*; D002455:Cell Division / Q000502:physiology; D002999:Clone Cells / Q000502:physiology; D003584:Cytological Techniques; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D005434:Flow Cytometry; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D016133:Polymerase Chain Reaction; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000302:isolation & purification; D051381:Rats; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D016159:Tumor Suppressor Protein p53