Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res 1991 Jan 01;51(1):365-71
Date
01/01/1991Pubmed ID
1703038Scopus ID
2-s2.0-0026079119 (requires institutional sign-in at Scopus site) 155 CitationsAbstract
Normal human esophageal autopsy tissue was explanted in serum-free medium. The epithelial outgrowths were subcultured and then transfected by strontium phosphate coprecipitation with plasmid pRSV-T consisting of the RSV-LTR promoter and the sequence encoding the simian virus 40 large T-antigen. The transfected cells, but not the sham-transfected controls, formed multilayered colonies within 3-4 weeks, after which the colonies were transferred and cell strains (HE-451 and HE-457) developed. Both cell strains grew exponentially for 8-10 weeks and then senesced. After a "crisis" of 6-8 months, growth resumed in isolated colonies. One line, HET-1A from HE-457, was developed and has now undergone more than 250 population doublings. This line has retained epithelial morphology, stains positively for cytokeratins and the simian virus 40 T-antigen gene by immunofluorescence, and has remained nontumorigenic in athymic, nude mice for more than 12 months. Karyotypic analysis by Giemsa banding has shown that HET-1A is hypodiploid (34-40 chromosomes). Growth factor studies have shown that HET-1A is stimulated by Ca2+, and inhibited by fetal bovine serum, transforming growth factor-beta 1, and transforming growth factor-beta 2. This serum-free immortalized esophageal cell system will be useful for investigating the action of putative esophageal carcinogens.
Author List
Stoner GD, Kaighn ME, Reddel RR, Resau JH, Bowman D, Naito Z, Matsukura N, You M, Galati AJ, Harris CCMESH terms used to index this publication - Major topics in bold
Antigens, Polyomavirus TransformingCalcium
Cell Division
Cell Line
Culture Media
DNA Fingerprinting
Epithelial Cells
Esophagus
Humans
In Vitro Techniques
Karyotyping
Keratins
Transfection
Transforming Growth Factors
Vimentin