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Cited by 607

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

Immortalization of Human Bronchial Epithelial Cells in the Absence of Viral Oncoproteins

Ramirez, Ruben D., Sheridan, Shelley, Girard, Luc, Sato, Mitsuo, Kim, Young, Pollack, Jon, Peyton, Michael, Zou, Ying, Kurie, Jonathan M., DiMaio, J. Michael, Milchgrub, Sara, Smith, Alice L., Souza, Rhonda F., Gilbey, Laura, Zhang, Xi, Gandia, Kenia, Vaughan, Melville B., Wright, Woodring E., Gazdar, Adi F., Shay, Jerry W., Minna, John D.·2004·Cancer Research
DOI 10.1158/0008-5472.can-04-3703

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Abstract

By expressing two genes (hTERT and Cdk4), we have developed a method to reproducibly generate continuously replicating human bronchial epithelial cell (HBEC) lines that provide a novel resource to study the molecular pathogenesis of lung cancer and the differentiation of bronchial epithelial cells. Twelve human bronchial epithelial biopsy specimens obtained from persons with and without lung cancer were placed into short-term culture and serially transfected with retroviral constructs containing cyclin-dependent kinase (Cdk) 4 and human telomerase reverse transcriptase (hTERT), resulting in continuously growing cultures. The order of introduction of Cdk4 and hTERT did not appear to be important; however, transfection of either gene alone did not result in immortalization. Although they could be cloned, the immortalized bronchial cells did not form colonies in soft agar or tumors in nude mice. The immortalized HBECs have epithelial morphology; express epithelial markers cytokeratins 7, 14, 17, and 19, the stem cell marker p63, and high levels of p16INK4a; and have an intact p53 checkpoint pathway. Cytogenetic analysis and array comparative genomic hybridization profiling show immortalized HBECs to have duplication of parts of chromosomes 5 and 20. Microarray gene expression profiling demonstrates that the Cdk4/hTERT-immortalized bronchial cell lines clustered together and with nonimmortalized bronchial cells, distinct from lung cancer cell lines. We also immortalized several parental cultures with viral oncoproteins human papilloma virus type 16 E6/E7 with and without hTERT, and these cells exhibited loss of the p53 checkpoint and significantly different gene expression profiles compared with Cdk4/hTERT-immortalized HBECs. These HBEC lines are a valuable new tool for studying of the pathogenesis of lung cancer.

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Crossref · 10.1158/0008-5472.can-04-3703

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Research works printed but not linked: 26

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Metadata

Authors
Ramirez, R. D., Sheridan, S., Girard, L., Sato, M., Kim, Y., Pollack, J., Peyton, M., Zou, Y., Kurie, J. M., DiMaio, J. M., Milchgrub, S., Smith, A. L., Souza, R. F., Gilbey, L., Zhang, X., Gandia, K., Vaughan, M. B., Wright, W. E., Gazdar, A. F., Shay, J. W., & Minna, J. D.
Affiliations
  • 1Hamon Center for Therapeutic Oncology Research and Departments of
  • 9Department of Pathology, Stanford University Medical Center, Stanford, California
  • 5Cell Biology, and
  • 8Department of Thoracic/Head and Neck Medical Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas; and
  • 6Cardiovascular and Thoracic Surgery, The University of Texas Southwestern Medical Center, Dallas, Texas;
  • 4Pathology,
  • 2Internal Medicine,
  • 7Dallas Veterans Affairs Medical Center, Dallas, Texas;
Venue
Cancer Research
Publisher
American Association for Cancer Research (AACR)
Published
2004-12-15
Volume
64
Issue
24
Pages
9027-9034
ISSN
  • 0008-5472
  • 1538-7445
DOI
10.1158/0008-5472.can-04-3703
Cited by
607
References
3
Language
en

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