cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer
<p>Abstract</p> <p>Background</p> <p>Recently published data showed discrepancies beteween <it>P53 </it>cDNA and DNA sequencing in glioblastomas. We hypothesised that similar discrepancies may be observed in other human cancers.</p> <p>Methods<...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2009-08-01
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| Series: | BMC Cancer |
| Online Access: | http://www.biomedcentral.com/1471-2407/9/278 |
| _version_ | 1818359998654185472 |
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| author | Jesionek-Kupnicka Dorota Stawski Robert Zawlik Izabela Kulczycka-Wojdala Dominika Pasz-Walczak Grazyna Rieske Piotr Zakrzewska Magdalena Szybka Malgorzata Liberski Pawel P Kordek Radzislaw |
| author_facet | Jesionek-Kupnicka Dorota Stawski Robert Zawlik Izabela Kulczycka-Wojdala Dominika Pasz-Walczak Grazyna Rieske Piotr Zakrzewska Magdalena Szybka Malgorzata Liberski Pawel P Kordek Radzislaw |
| author_sort | Jesionek-Kupnicka Dorota |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Recently published data showed discrepancies beteween <it>P53 </it>cDNA and DNA sequencing in glioblastomas. We hypothesised that similar discrepancies may be observed in other human cancers.</p> <p>Methods</p> <p>To this end, we analyzed 23 colorectal cancers for <it>P53 </it>mutations and gene expression using both DNA and cDNA sequencing, real-time PCR and immunohistochemistry.</p> <p>Results</p> <p>We found <it>P53 </it>gene mutations in 16 cases (15 missense and 1 nonsense). Two of the 15 cases with missense mutations showed alterations based only on cDNA, and not DNA sequencing. Moreover, in 6 of the 15 cases with a cDNA mutation those mutations were difficult to detect in the DNA sequencing, so the results of DNA analysis alone could be misinterpreted if the cDNA sequencing results had not also been available. In all those 15 cases, we observed a higher ratio of the mutated to the wild type template by cDNA analysis, but not by the DNA analysis. Interestingly, a similar overexpression of <it>P53 </it>mRNA was present in samples with and without <it>P53 </it>mutations.</p> <p>Conclusion</p> <p>In terms of colorectal cancer, those discrepancies might be explained under three conditions: 1, overexpression of mutated <it>P53 </it>mRNA in cancer cells as compared with normal cells; 2, a higher content of cells without <it>P53 </it>mutation (normal cells and cells showing <it>K-RAS </it>and/or <it>APC </it>but not <it>P53 </it>mutation) in samples presenting <it>P53 </it>mutation; 3, heterozygous or hemizygous mutations of <it>P53 </it>gene. Additionally, for heterozygous mutations unknown mechanism(s) causing selective overproduction of mutated allele should also be considered. Our data offer new clues for studying discrepancy in <it>P53 </it>cDNA and DNA sequencing analysis.</p> |
| first_indexed | 2024-12-13T20:53:48Z |
| format | Article |
| id | doaj.art-c6cca4ad51f7482a93a888bd4699842a |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-12-13T20:53:48Z |
| publishDate | 2009-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-c6cca4ad51f7482a93a888bd4699842a2022-12-21T23:31:48ZengBMCBMC Cancer1471-24072009-08-019127810.1186/1471-2407-9-278cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancerJesionek-Kupnicka DorotaStawski RobertZawlik IzabelaKulczycka-Wojdala DominikaPasz-Walczak GrazynaRieske PiotrZakrzewska MagdalenaSzybka MalgorzataLiberski Pawel PKordek Radzislaw<p>Abstract</p> <p>Background</p> <p>Recently published data showed discrepancies beteween <it>P53 </it>cDNA and DNA sequencing in glioblastomas. We hypothesised that similar discrepancies may be observed in other human cancers.</p> <p>Methods</p> <p>To this end, we analyzed 23 colorectal cancers for <it>P53 </it>mutations and gene expression using both DNA and cDNA sequencing, real-time PCR and immunohistochemistry.</p> <p>Results</p> <p>We found <it>P53 </it>gene mutations in 16 cases (15 missense and 1 nonsense). Two of the 15 cases with missense mutations showed alterations based only on cDNA, and not DNA sequencing. Moreover, in 6 of the 15 cases with a cDNA mutation those mutations were difficult to detect in the DNA sequencing, so the results of DNA analysis alone could be misinterpreted if the cDNA sequencing results had not also been available. In all those 15 cases, we observed a higher ratio of the mutated to the wild type template by cDNA analysis, but not by the DNA analysis. Interestingly, a similar overexpression of <it>P53 </it>mRNA was present in samples with and without <it>P53 </it>mutations.</p> <p>Conclusion</p> <p>In terms of colorectal cancer, those discrepancies might be explained under three conditions: 1, overexpression of mutated <it>P53 </it>mRNA in cancer cells as compared with normal cells; 2, a higher content of cells without <it>P53 </it>mutation (normal cells and cells showing <it>K-RAS </it>and/or <it>APC </it>but not <it>P53 </it>mutation) in samples presenting <it>P53 </it>mutation; 3, heterozygous or hemizygous mutations of <it>P53 </it>gene. Additionally, for heterozygous mutations unknown mechanism(s) causing selective overproduction of mutated allele should also be considered. Our data offer new clues for studying discrepancy in <it>P53 </it>cDNA and DNA sequencing analysis.</p>http://www.biomedcentral.com/1471-2407/9/278 |
| spellingShingle | Jesionek-Kupnicka Dorota Stawski Robert Zawlik Izabela Kulczycka-Wojdala Dominika Pasz-Walczak Grazyna Rieske Piotr Zakrzewska Magdalena Szybka Malgorzata Liberski Pawel P Kordek Radzislaw cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer BMC Cancer |
| title | cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer |
| title_full | cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer |
| title_fullStr | cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer |
| title_full_unstemmed | cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer |
| title_short | cDNA sequencing improves the detection of <it>P53 </it>missense mutations in colorectal cancer |
| title_sort | cdna sequencing improves the detection of it p53 it missense mutations in colorectal cancer |
| url | http://www.biomedcentral.com/1471-2407/9/278 |
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