Mutational analysis of the p21/WAF1/CIP1/SDI1 coding region in human tumor cell lines

Terry, L. A., Boyd, J., Alcorta, D., Lyon, T., Solomon, G., Hannon, G., Berchuck, A., Beach, D., Barrett, J. C. (1996) Mutational analysis of the p21/WAF1/CIP1/SDI1 coding region in human tumor cell lines. Molecular Carcinogenesis, 16 (4). pp. 221-228. ISSN 08991987 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/8784465
DOI: 10.1002/(sici)1098-2744(199608)16:4<221::aid-mc6>3.0.co;2-i

Abstract

p21/WAF1/CIP1/SDI1 is an important cell-cycle mediator with tumor suppressor gene capabilities, and its inactivation could potentially lead to tumor progression. Because tumor suppressor genes are commonly inactivated by somatic and germline mutations, we analyzed a variety of human tumor cell lines for p21 mutations. We used single-strand conformational analysis and direct sequencing to identify possible mutations in the p21 coding region. Two base-alterations were observed in 41 immortalized human tumor cell lines. A previously reported polymorphism that results in a serine-to-arginine amino-acid substitution at codon 31 was found in 24% (10 of 41) of the tumor cell lines but was also found in 10% (six of 62) of normal parental DNAs tested and 7% (three of 43) of normal DNAs from patients with primary endometrial tumors. Another nucleotide substitution found at codon 80 resulted in the replacement of threonine with methionine. Codon 80 changes were found in 7% (three of 41) of the tumor cell lines (all endometrial) and in 2% (one of 62) of the normal parental DNAs. This change was not found in any of the primary endometrial tumors examined. The biological activity of these base changes was analyzed by using in vitro cyclin-dependent kinase 2-cyclin A kinase assays and calcium phosphate transfections. We observed that wild-type p21 and the p21 variants had similar growth-inhibitory abilities. Thus, our results suggest that mutation of the p21 gene is not prevalent in human tumor cell lines and is not a probable mechanism of inactivation of this gene.

Item Type: Paper
Uncontrolled Keywords: Cell-cycle regulation Cyclin-dependent kinase inhibitor p21 Polymorphism Single-strand conformational analysis cyclin dependent kinase inhibitor amino acid substitution article cell cycle controlled study gene mutation human human cell priority journal tumor cell line tumor suppressor gene Base Sequence Cell Division Cyclin-Dependent Kinase Inhibitor p21 Cyclins DNA Mutational Analysis DNA, Neoplasm Humans Molecular Sequence Data Mutation Neoplasms Polymorphism, Genetic Tumor Cells, Cultured
Subjects: diseases & disorders > neoplasms
organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > cell cycle
organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > cell division
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations
CSHL Authors:
Communities: CSHL labs > Beach lab
CSHL labs > Hannon lab
Depositing User: Kathleen Darby
Date Deposited: 13 May 2014 14:37
Last Modified: 13 May 2014 14:37
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30117

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