Multiple oncogenic changes (K-RASV12, p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells

Mitsuo Sato, Melville B. Vaughan, Luc Girard, Michael Peyton, Woochang Lee, David S. Shames, Ruben D. Ramirez, Noriaki Sunaga, Adi F. Gazdar, Jerry W. Shay, John D. Minna

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194 Citations (Scopus)

Abstract

We evaluated the contribution of three genetic alterations (p53 knockdown, K-RASV12, and mutant EGFR) to lung tumorigenesis using human bronchial epithelial cells (HBEC) immortalized with telomerase and Cdk4-mediated p16 bypass. RNA interference p53 knockdown or oncogenic K-RASV12 resulted in enhanced anchorage-independent growth and increased saturation density of HBECs. The combination of p53 knockdown and K-RASV12 further enhanced the tumorigenic phenotype with increased growth in soft agar and an invasive phenotype in three-dimensional organotypic cultures but failed to cause HBECs to form tumors in nude mice. Growth of HBECs was highly dependent on epidermal growth factor (EGF) and completely inhibited by EGF receptor (EGFR) tyrosine kinase inhibitors, which induced G1 arrest. Introduction of EGFR mutations E746-A750 del and L858R progressed HBECs toward malignancy as measured by soft agar growth, including EGF-independent growth, but failed to induce tumor formation. Mutant EGFRs were associated with higher levels of phospho-Akt, phospho-signal transducers and activators of transcription 3 [but not phospho-extracellular signal-regulated kinase (ERK) 1/2], and increased expression of DUSP6/MKP-3 phosphatase (an inhibitor of phospho-ERK1/2). These results indicate that (a) the HBEC model system is a powerful new approach to assess the contribution of individual and combinations of genetic alterations to lung cancer pathogenesis; (b) a combination of four genetic alterations, including human telomerase reverse transcriptase overexpression, bypass of p16/RB and p53 pathways, and mutant K-RAS V12 or mutant EGFR, is still not sufficient for HBECs to completely transform to cancer; and (c) EGFR tyrosine kinase inhibitors inhibit the growth of preneoplastic HBEC cells, suggesting their potential for chemoprevention.

Original languageEnglish (US)
Pages (from-to)2116-2128
Number of pages13
JournalCancer Research
Volume66
Issue number4
DOIs
StatePublished - Feb 15 2006

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Telomerase
Epidermal Growth Factor Receptor
Epithelial Cells
Phenotype
Growth
Epidermal Growth Factor
Protein-Tyrosine Kinases
Agar
Dual Specificity Phosphatase 6
Neoplasms
STAT3 Transcription Factor
Growth Inhibitors
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Chemoprevention
RNA Interference
Nude Mice
Lung Neoplasms
Carcinogenesis
Lung

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Multiple oncogenic changes (K-RASV12, p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells. / Sato, Mitsuo; Vaughan, Melville B.; Girard, Luc; Peyton, Michael; Lee, Woochang; Shames, David S.; Ramirez, Ruben D.; Sunaga, Noriaki; Gazdar, Adi F.; Shay, Jerry W.; Minna, John D.

In: Cancer Research, Vol. 66, No. 4, 15.02.2006, p. 2116-2128.

Research output: Contribution to journalArticle

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abstract = "We evaluated the contribution of three genetic alterations (p53 knockdown, K-RASV12, and mutant EGFR) to lung tumorigenesis using human bronchial epithelial cells (HBEC) immortalized with telomerase and Cdk4-mediated p16 bypass. RNA interference p53 knockdown or oncogenic K-RASV12 resulted in enhanced anchorage-independent growth and increased saturation density of HBECs. The combination of p53 knockdown and K-RASV12 further enhanced the tumorigenic phenotype with increased growth in soft agar and an invasive phenotype in three-dimensional organotypic cultures but failed to cause HBECs to form tumors in nude mice. Growth of HBECs was highly dependent on epidermal growth factor (EGF) and completely inhibited by EGF receptor (EGFR) tyrosine kinase inhibitors, which induced G1 arrest. Introduction of EGFR mutations E746-A750 del and L858R progressed HBECs toward malignancy as measured by soft agar growth, including EGF-independent growth, but failed to induce tumor formation. Mutant EGFRs were associated with higher levels of phospho-Akt, phospho-signal transducers and activators of transcription 3 [but not phospho-extracellular signal-regulated kinase (ERK) 1/2], and increased expression of DUSP6/MKP-3 phosphatase (an inhibitor of phospho-ERK1/2). These results indicate that (a) the HBEC model system is a powerful new approach to assess the contribution of individual and combinations of genetic alterations to lung cancer pathogenesis; (b) a combination of four genetic alterations, including human telomerase reverse transcriptase overexpression, bypass of p16/RB and p53 pathways, and mutant K-RAS V12 or mutant EGFR, is still not sufficient for HBECs to completely transform to cancer; and (c) EGFR tyrosine kinase inhibitors inhibit the growth of preneoplastic HBEC cells, suggesting their potential for chemoprevention.",
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AU - Sato, Mitsuo

AU - Vaughan, Melville B.

AU - Girard, Luc

AU - Peyton, Michael

AU - Lee, Woochang

AU - Shames, David S.

AU - Ramirez, Ruben D.

AU - Sunaga, Noriaki

AU - Gazdar, Adi F.

AU - Shay, Jerry W.

AU - Minna, John D.

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