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. 1993 Jul;119(9):522–526. doi: 10.1007/BF01686461

Alteration in the retinoblastoma gene associated with immortalization of human fibroblasts treated with60Co gamma rays

Akira Endo 1,✉, Yoshio Kano 1, Koichiro Mihara 1, Kunzo Orita 2, Masayoshi Namba 1
PMCID: PMC12201876  PMID: 8100822

Abstract

Genetic analysis was carried out in human fibroblasts (KMST-6) immortalized by treatment with60Co gamma rays in order to determine if any genetic change was involved in the immortal transformation of human cells. Analysis by restriction fragment length polymorphism revealed an alteration in chromosome 13q12–14, in which the retinoblastoma (RB) gene locus (13q14) is located. Then the RB gene itself was examined. Structural abnormalities in the RB gene were detected by Southern blot analysis. Furthermore, abnormal RB protein (pRB) was expressed in immortalized KMST-6 cells, as shown by in vitro phosphorylation, whereas normal KMS-6 cells expressed the intact pRB. These findings indicated that inactivation of the RB gene is one of the key events of the immortalization of human cells.

Key words: Cell transformation, Fibroblasts, Retinoblastoma gene

Abbreviations

RB

retinoblastoma

pRB

retinoblastoma gene product (protein)

T

simian virus 40 large T antigen

E1A

adenovirus E1A protein

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