Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.

A comprehensive analysis of the TP53 gene and its protein status was carried out on a panel of 56 colorectal cancer cell lines. This analysis was based on a combination of denaturing HPLC mutation screening of all exons of the p53 gene, sequencing the cDNA, and assessing the function of the p53 prot...

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Main Authors: Liu, Y, Bodmer, W
Format: Journal article
Language:English
Published: 2006
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author Liu, Y
Bodmer, W
author_facet Liu, Y
Bodmer, W
author_sort Liu, Y
collection OXFORD
description A comprehensive analysis of the TP53 gene and its protein status was carried out on a panel of 56 colorectal cancer cell lines. This analysis was based on a combination of denaturing HPLC mutation screening of all exons of the p53 gene, sequencing the cDNA, and assessing the function of the p53 protein by assaying the induced expression of phosphorylated p53 and p21 after exposing cells to gamma-rays. In a few cases where there was no production of p53 message nor evidence of functional p53 protein, all of the p53 exons were sequenced directly. Thirteen of the 56 cell lines had functional p53, 21 lines had missense mutations (one of which made no detectable protein), 4 lines produced no p53 transcripts, and the remaining 18 lines carried truncating TP53 mutations. Thus, our results showed a relatively high frequency of TP53 mutations (76.8%) in our cell lines, with almost half of the mutations being truncating mutations. This is a rather higher frequency of such mutations than usually reported. Of the 18 cell lines with truncating mutations, 12 had detectable truncated protein based on Western blot analysis, whereas no protein was detected in the remaining 6 cell lines. Our data provide a valuable source of TP 53 mutations for further studies and raise the question of the extent to which truncating mutations may have dominant negative effects, even when no truncated protein can be detected by standard methods.
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spelling oxford-uuid:a19c05ed-1960-4c2b-b8e1-fb90013940ab2022-03-27T02:14:22ZAnalysis of P53 mutations and their expression in 56 colorectal cancer cell lines.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a19c05ed-1960-4c2b-b8e1-fb90013940abEnglishSymplectic Elements at Oxford2006Liu, YBodmer, WA comprehensive analysis of the TP53 gene and its protein status was carried out on a panel of 56 colorectal cancer cell lines. This analysis was based on a combination of denaturing HPLC mutation screening of all exons of the p53 gene, sequencing the cDNA, and assessing the function of the p53 protein by assaying the induced expression of phosphorylated p53 and p21 after exposing cells to gamma-rays. In a few cases where there was no production of p53 message nor evidence of functional p53 protein, all of the p53 exons were sequenced directly. Thirteen of the 56 cell lines had functional p53, 21 lines had missense mutations (one of which made no detectable protein), 4 lines produced no p53 transcripts, and the remaining 18 lines carried truncating TP53 mutations. Thus, our results showed a relatively high frequency of TP53 mutations (76.8%) in our cell lines, with almost half of the mutations being truncating mutations. This is a rather higher frequency of such mutations than usually reported. Of the 18 cell lines with truncating mutations, 12 had detectable truncated protein based on Western blot analysis, whereas no protein was detected in the remaining 6 cell lines. Our data provide a valuable source of TP 53 mutations for further studies and raise the question of the extent to which truncating mutations may have dominant negative effects, even when no truncated protein can be detected by standard methods.
spellingShingle Liu, Y
Bodmer, W
Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title_full Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title_fullStr Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title_full_unstemmed Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title_short Analysis of P53 mutations and their expression in 56 colorectal cancer cell lines.
title_sort analysis of p53 mutations and their expression in 56 colorectal cancer cell lines
work_keys_str_mv AT liuy analysisofp53mutationsandtheirexpressionin56colorectalcancercelllines
AT bodmerw analysisofp53mutationsandtheirexpressionin56colorectalcancercelllines