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

Cell Biosci 2016 11;6:57. Epub 2016 Nov 11.

Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, NIH., Bethesda, MD USA.

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 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 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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http://dx.doi.org/10.1186/s13578-016-0122-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106784PMC
November 2016
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References

(Supplied by CrossRef)

JM Sedivy et al.
Proc Natl Acad Sci USA 1998

RD Ramirez et al.
Genes Dev 2001

B Vogelstein et al.
Science 2013

Y Katakura et al.
Methods Cell Biol 1998

D Hanahan et al.
Cell 2000

WC Hahn et al.
N Engl J Med 2002

DS Neufeld et al.
Mol Cell Biol 1987

JW Shay et al.
Biochim Biophys Acta 1991

K M√ľnger et al.
J Virol 1989

P Hawley-Nelson et al.
EMBO J 1989

S Watanabe et al.
J Virol 1989

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