title: Immortalization and transformation of primary human airway epithelial cells by gene transfer.
abstract: One critical step in the development of a cancerous cell is its acquisition of an unlimited replicative lifespan, the process termed immortalization. Experimental model systems designed to study cellular transformation ex vivo have relied to date on the in vitro selection of a subpopulation of cells that have become immortalized through treatment with chemical or physical mutagens and the selection of rare clonal variants. In this study, we describe the direct immortalization of primary human airway epithelial cells through the successive introduction of the Simian Virus 40 Early Region and the telomerase catalytic subunit hTERT. Cells immortalized in this way are now responsive to malignant transformation by an introduced H-ras or K-ras oncogene. These immortalized human airway epithelial cells, which have been created through the stepwise introduction of genetic alterations, provide a novel experimental model system with which to study further the biology of the airway epithelial cell and to dissect the molecular basis of lung cancer pathogenesis.
authors.affiliation: Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Ante S
authors.initials: AS
authors.lastname: Lundberg
authors.forename: Scott H
authors.initials: SH
authors.lastname: Randell
authors.forename: Sheila A
authors.initials: SA
authors.lastname: Stewart
authors.forename: Brian
authors.initials: B
authors.lastname: Elenbaas
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Hartwell
authors.forename: Mary W
authors.initials: MW
authors.lastname: Brooks
authors.forename: Mark D
authors.initials: MD
authors.lastname: Fleming
authors.forename: John C
authors.initials: JC
authors.lastname: Olsen
authors.forename: Scott W
authors.initials: SW
authors.lastname: Miller
authors.forename: Robert A
authors.initials: RA
authors.lastname: Weinberg
authors.forename: William C
authors.initials: WC
authors.lastname: Hahn
mesh_terms: D015153:Blotting, Western; D001980:Bronchi / Q000378:metabolism* / Q000473:pathology*; D002454:Cell Differentiation; D002455:Cell Division; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic / Q000235:genetics* / Q000473:pathology*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology*; D011905:Genes, ras / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D013539:Simian virus 40 / Q000235:genetics; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic
chemical_list: D004268:DNA-Binding Proteins; D019098:Telomerase