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Reliable blood cancer cells' telomere length evaluation by qPCR

BACKGROUND: Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost‐effective approach to study samples...

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Autores principales: Ropio, Joana, Chebly, Alain, Ferrer, Jacky, Prochazkova‐Carlotti, Martina, Idrissi, Yamina, Azzi‐Martin, Lamia, Cappellen, David, Pham‐Ledard, Anne, Soares, Paula, Merlio, Jean‐Philippe, Chevret, Edith
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7196062/
https://www.ncbi.nlm.nih.gov/pubmed/32142223
http://dx.doi.org/10.1002/cam4.2816
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author Ropio, Joana
Chebly, Alain
Ferrer, Jacky
Prochazkova‐Carlotti, Martina
Idrissi, Yamina
Azzi‐Martin, Lamia
Cappellen, David
Pham‐Ledard, Anne
Soares, Paula
Merlio, Jean‐Philippe
Chevret, Edith
author_facet Ropio, Joana
Chebly, Alain
Ferrer, Jacky
Prochazkova‐Carlotti, Martina
Idrissi, Yamina
Azzi‐Martin, Lamia
Cappellen, David
Pham‐Ledard, Anne
Soares, Paula
Merlio, Jean‐Philippe
Chevret, Edith
author_sort Ropio, Joana
collection PubMed
description BACKGROUND: Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost‐effective approach to study samples with low DNA amounts. METHODS: Cancer cells’ telomere length was evaluated by relative and absolute qPCR methods. RESULTS: Robust and reproducible telomere length measurements were optimized taking into account a careful reference gene selection and by knowing the cancer cells ploidy. qPCR data were compared to “gold standard” measurement from terminal restriction fragment (TRF). CONCLUSIONS: Our study provides guidance and recommendations for accurate telomere length measurement by qPCR in cancer cells, taking advantage of our expertise in telomere homeostasis investigation in primary cutaneous T‐cell lymphomas. Furthermore, our data emphasize the requirement of samples with both, high DNA quality and high tumor cells representation.
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spelling pubmed-71960622020-05-04 Reliable blood cancer cells' telomere length evaluation by qPCR Ropio, Joana Chebly, Alain Ferrer, Jacky Prochazkova‐Carlotti, Martina Idrissi, Yamina Azzi‐Martin, Lamia Cappellen, David Pham‐Ledard, Anne Soares, Paula Merlio, Jean‐Philippe Chevret, Edith Cancer Med Cancer Biology BACKGROUND: Telomere shortening is linked to a range of different human diseases, hence reliable measurement methods are needed to uncover such associations. Among the plethora of telomere length measurement methods, qPCR is reported as easy to conduct and a cost‐effective approach to study samples with low DNA amounts. METHODS: Cancer cells’ telomere length was evaluated by relative and absolute qPCR methods. RESULTS: Robust and reproducible telomere length measurements were optimized taking into account a careful reference gene selection and by knowing the cancer cells ploidy. qPCR data were compared to “gold standard” measurement from terminal restriction fragment (TRF). CONCLUSIONS: Our study provides guidance and recommendations for accurate telomere length measurement by qPCR in cancer cells, taking advantage of our expertise in telomere homeostasis investigation in primary cutaneous T‐cell lymphomas. Furthermore, our data emphasize the requirement of samples with both, high DNA quality and high tumor cells representation. John Wiley and Sons Inc. 2020-03-06 /pmc/articles/PMC7196062/ /pubmed/32142223 http://dx.doi.org/10.1002/cam4.2816 Text en © 2020 The Authors. Cancer Medicine published by John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Cancer Biology
Ropio, Joana
Chebly, Alain
Ferrer, Jacky
Prochazkova‐Carlotti, Martina
Idrissi, Yamina
Azzi‐Martin, Lamia
Cappellen, David
Pham‐Ledard, Anne
Soares, Paula
Merlio, Jean‐Philippe
Chevret, Edith
Reliable blood cancer cells' telomere length evaluation by qPCR
title Reliable blood cancer cells' telomere length evaluation by qPCR
title_full Reliable blood cancer cells' telomere length evaluation by qPCR
title_fullStr Reliable blood cancer cells' telomere length evaluation by qPCR
title_full_unstemmed Reliable blood cancer cells' telomere length evaluation by qPCR
title_short Reliable blood cancer cells' telomere length evaluation by qPCR
title_sort reliable blood cancer cells' telomere length evaluation by qpcr
topic Cancer Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7196062/
https://www.ncbi.nlm.nih.gov/pubmed/32142223
http://dx.doi.org/10.1002/cam4.2816
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