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Limited importance of the dominant-negative effect of TP53 missense mutations
BACKGROUND: Heterozygosity of TP53 missense mutations is related to the phenomenon of the dominant-negative effect (DNE). To estimate the importance of the DNE of TP53 mutations, we analysed the percentage of cancer cases showing a single heterozygous mutation of TP53 and searched for a cell line wi...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3129589/ https://www.ncbi.nlm.nih.gov/pubmed/21668955 http://dx.doi.org/10.1186/1471-2407-11-243 |
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author | Stoczynska-Fidelus, Ewelina Szybka, Malgorzata Piaskowski, Sylwester Bienkowski, Michal Hulas-Bigoszewska, Krystyna Banaszczyk, Mateusz Zawlik, Izabela Jesionek-Kupnicka, Dorota Kordek, Radzislaw Liberski, Pawel P Rieske, Piotr |
author_facet | Stoczynska-Fidelus, Ewelina Szybka, Malgorzata Piaskowski, Sylwester Bienkowski, Michal Hulas-Bigoszewska, Krystyna Banaszczyk, Mateusz Zawlik, Izabela Jesionek-Kupnicka, Dorota Kordek, Radzislaw Liberski, Pawel P Rieske, Piotr |
author_sort | Stoczynska-Fidelus, Ewelina |
collection | PubMed |
description | BACKGROUND: Heterozygosity of TP53 missense mutations is related to the phenomenon of the dominant-negative effect (DNE). To estimate the importance of the DNE of TP53 mutations, we analysed the percentage of cancer cases showing a single heterozygous mutation of TP53 and searched for a cell line with a single heterozygous mutation of this gene. This approach was based on the knowledge that genes with evident DNE, such as EGFR and IDH1, represent nearly 100% of single heterozygous mutations in tumour specimens and cell lines. METHODS: Genetic analyses (LOH and sequencing) performed for early and late passages of several cell lines originally described as showing single heterozygous TP53 mutations (H-318, G-16, PF-382, MOLT-13, ST-486 and LS-123). Statistical analysis of IARC TP53 and SANGER databases. Genetic analyses of N-RAS, FBXW7, PTEN and STR markers to test cross-contamination and cell line identity. Cell cloning, fluorescence-activated cell sorting and SSCP performed for the PF-382 cell line. RESULTS: A database study revealed TP53 single heterozygous mutations in 35% of in vivo (surgical and biopsy) samples and only 10% of cultured cells (in vitro), although those numbers appeared to be overestimated. We deem that published in vivo TP53 mutation analyses are not as rigorous as studies in vitro, and we did not find any cell line showing a stable, single heterozygous mutation. G16, PF-382 and MOLT-13 cells harboured single heterozygous mutations temporarily. ST-486, H-318 and LS-123 cell lines were misclassified. Specific mutations, such as R175H, R273H, R273L or R273P, which are reported in the literature to exert a DNE, showed the lowest percentage of single heterozygous mutations in vitro (about 5%). CONCLUSION: We suggest that the currently reported percentage of TP53 single heterozygous mutations in tumour samples and cancer cell lines is overestimated. Thus, the magnitude of the DNE of TP53 mutations is questionable. This scepticism is supported by database investigations showing that retention of the wild-type allele occurs with the same frequency as either nonsense or missense TP53 mutations. |
format | Online Article Text |
id | pubmed-3129589 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-31295892011-07-06 Limited importance of the dominant-negative effect of TP53 missense mutations Stoczynska-Fidelus, Ewelina Szybka, Malgorzata Piaskowski, Sylwester Bienkowski, Michal Hulas-Bigoszewska, Krystyna Banaszczyk, Mateusz Zawlik, Izabela Jesionek-Kupnicka, Dorota Kordek, Radzislaw Liberski, Pawel P Rieske, Piotr BMC Cancer Research Article BACKGROUND: Heterozygosity of TP53 missense mutations is related to the phenomenon of the dominant-negative effect (DNE). To estimate the importance of the DNE of TP53 mutations, we analysed the percentage of cancer cases showing a single heterozygous mutation of TP53 and searched for a cell line with a single heterozygous mutation of this gene. This approach was based on the knowledge that genes with evident DNE, such as EGFR and IDH1, represent nearly 100% of single heterozygous mutations in tumour specimens and cell lines. METHODS: Genetic analyses (LOH and sequencing) performed for early and late passages of several cell lines originally described as showing single heterozygous TP53 mutations (H-318, G-16, PF-382, MOLT-13, ST-486 and LS-123). Statistical analysis of IARC TP53 and SANGER databases. Genetic analyses of N-RAS, FBXW7, PTEN and STR markers to test cross-contamination and cell line identity. Cell cloning, fluorescence-activated cell sorting and SSCP performed for the PF-382 cell line. RESULTS: A database study revealed TP53 single heterozygous mutations in 35% of in vivo (surgical and biopsy) samples and only 10% of cultured cells (in vitro), although those numbers appeared to be overestimated. We deem that published in vivo TP53 mutation analyses are not as rigorous as studies in vitro, and we did not find any cell line showing a stable, single heterozygous mutation. G16, PF-382 and MOLT-13 cells harboured single heterozygous mutations temporarily. ST-486, H-318 and LS-123 cell lines were misclassified. Specific mutations, such as R175H, R273H, R273L or R273P, which are reported in the literature to exert a DNE, showed the lowest percentage of single heterozygous mutations in vitro (about 5%). CONCLUSION: We suggest that the currently reported percentage of TP53 single heterozygous mutations in tumour samples and cancer cell lines is overestimated. Thus, the magnitude of the DNE of TP53 mutations is questionable. This scepticism is supported by database investigations showing that retention of the wild-type allele occurs with the same frequency as either nonsense or missense TP53 mutations. BioMed Central 2011-06-13 /pmc/articles/PMC3129589/ /pubmed/21668955 http://dx.doi.org/10.1186/1471-2407-11-243 Text en Copyright ©2011 Stoczynska-Fidelus et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Stoczynska-Fidelus, Ewelina Szybka, Malgorzata Piaskowski, Sylwester Bienkowski, Michal Hulas-Bigoszewska, Krystyna Banaszczyk, Mateusz Zawlik, Izabela Jesionek-Kupnicka, Dorota Kordek, Radzislaw Liberski, Pawel P Rieske, Piotr Limited importance of the dominant-negative effect of TP53 missense mutations |
title | Limited importance of the dominant-negative effect of TP53 missense mutations |
title_full | Limited importance of the dominant-negative effect of TP53 missense mutations |
title_fullStr | Limited importance of the dominant-negative effect of TP53 missense mutations |
title_full_unstemmed | Limited importance of the dominant-negative effect of TP53 missense mutations |
title_short | Limited importance of the dominant-negative effect of TP53 missense mutations |
title_sort | limited importance of the dominant-negative effect of tp53 missense mutations |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3129589/ https://www.ncbi.nlm.nih.gov/pubmed/21668955 http://dx.doi.org/10.1186/1471-2407-11-243 |
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