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One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation

BACKGROUND: The ability to transform normal human cells into cancer cells with the introduction of defined genetic alterations is a valuable method for understanding the mechanisms of oncogenesis. Easy establishment of immortalized but non-transformed human cells from various tissues would facilitat...

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Autores principales: Smith, Jordan L., Lee, Liam C., Read, Abigail, Li, Qiuning, Yu, Bing, Lee, Chih-Shia, Luo, Ji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106784/
https://www.ncbi.nlm.nih.gov/pubmed/27891214
http://dx.doi.org/10.1186/s13578-016-0122-6
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author Smith, Jordan L.
Lee, Liam C.
Read, Abigail
Li, Qiuning
Yu, Bing
Lee, Chih-Shia
Luo, Ji
author_facet Smith, Jordan L.
Lee, Liam C.
Read, Abigail
Li, Qiuning
Yu, Bing
Lee, Chih-Shia
Luo, Ji
author_sort Smith, Jordan L.
collection PubMed
description BACKGROUND: The ability to transform normal human cells into cancer cells with the introduction of defined genetic alterations is a valuable method for understanding the mechanisms of oncogenesis. Easy establishment of immortalized but non-transformed human cells from various tissues would facilitate these genetic analyses. RESULTS: We report here a simple, one-step immortalization method that involves retroviral vector mediated co-expression of the human telomerase protein and a shRNA targeting the CDKN2A gene locus. We demonstrate that this method could successfully immortalize human small airway epithelial cells while maintaining their chromosomal stability. We further showed that these cells retain p53 activity and can be transformed by the KRAS oncogene. CONCLUSIONS: Our method simplifies the immortalization process and is broadly applicable for establishing immortalized epithelial cell lines from primary human tissues for cancer research.
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spelling pubmed-51067842016-11-25 One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation Smith, Jordan L. Lee, Liam C. Read, Abigail Li, Qiuning Yu, Bing Lee, Chih-Shia Luo, Ji Cell Biosci Methodology BACKGROUND: The ability to transform normal human cells into cancer cells with the introduction of defined genetic alterations is a valuable method for understanding the mechanisms of oncogenesis. Easy establishment of immortalized but non-transformed human cells from various tissues would facilitate these genetic analyses. RESULTS: We report here a simple, one-step immortalization method that involves retroviral vector mediated co-expression of the human telomerase protein and a shRNA targeting the CDKN2A gene locus. We demonstrate that this method could successfully immortalize human small airway epithelial cells while maintaining their chromosomal stability. We further showed that these cells retain p53 activity and can be transformed by the KRAS oncogene. CONCLUSIONS: Our method simplifies the immortalization process and is broadly applicable for establishing immortalized epithelial cell lines from primary human tissues for cancer research. BioMed Central 2016-11-11 /pmc/articles/PMC5106784/ /pubmed/27891214 http://dx.doi.org/10.1186/s13578-016-0122-6 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Smith, Jordan L.
Lee, Liam C.
Read, Abigail
Li, Qiuning
Yu, Bing
Lee, Chih-Shia
Luo, Ji
One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title_full One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title_fullStr One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title_full_unstemmed One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title_short One-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
title_sort one-step immortalization of primary human airway epithelial cells capable of oncogenic transformation
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106784/
https://www.ncbi.nlm.nih.gov/pubmed/27891214
http://dx.doi.org/10.1186/s13578-016-0122-6
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