The p21WAF1/CIP1 promoter is methylated in rat-1 cells:: Stable restoration of p53-dependent p21WAF1/CIP1 expression after transfection of a genomic clone containing the p21WAF1/CIP1 gene

被引：64

作者：

Allan, LA ^{[1
]}

Duhig, T ^{[1
]}

Read, M ^{[1
]}

Fried, M ^{[1
]}

机构：

[1] Imperial Canc Res Fund, Eukaryot Gene Org & Express Lab, London WC2A 3PX, England

来源：

MOLECULAR AND CELLULAR BIOLOGY | 2000年 / 20卷 / 04期

关键词：

D O I：

10.1128/MCB.20.4.1291-1298.2000

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Rat-1 cells are used in many studies on transformation, cell cycle, and apoptosis. whereas UV treatment of Rat-1 cells results in apoptosis. X-ray treatment does not induce either apoptosis or a cell cycle block. X-ray treatment of Rat-1 cells results in both an increase of p53 protein and expression of the p53-inducible gene MDM2 but not the protein or mRNA of the p53-inducible p21(WAF1/CIP1) gene, which in other cells plays an important role in p53-mediated cell cycle block. The lack of p21(WAF1/CIP1) expression appears to be the result of hypermethylation of the p21(WAF1/CIP1) promoter region, as p21(WAF1/CIP1) protein expression could be induced by growth of Rat-1 cells in the presence of 5-aza-2-deoxycytidine. Furthermore, sequence analysis of bisulfite-treated DNA demonstrated extensive methylation of cytosine residues in CpG dinucleotides in a CpG-rich island in the promoter region of the p21(WAF1/CIP1) gene. Stable X-ray-induced p53-dependent p21(WAF1/CIP1) expression and cell cycle block were restored to a Rat-1 clone after transfection with a P1 artificial chromosome (PAC) DNA clone containing a rat genomic copy of the p21(WAF1/CIP1) gene. The absence of expression of the p21(WAF1/CIP1) gene may contribute to the suitability of Rat-1 cells for transformation, cell cycle, and apoptosis studies.

引用

页码：1291 / 1298

页数：8

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