Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells.
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.
Stoner
G D
GD
Department of Pathology, Medical College of Ohio, Toledo 43699.
Kaighn
M E
ME
Reddel
R R
RR
Resau
J H
JH
Bowman
D
D
Naito
Z
Z
Matsukura
N
N
You
M
M
Galati
A J
AJ
Harris
C C
CC
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D002118:Calcium / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media; D016172:DNA Fingerprinting; D004847:Epithelial Cells; D004947:Esophagus / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D007621:Karyotyping; D007633:Keratins / Q000378:metabolism; D014162:Transfection; D015291:Transforming Growth Factors / Q000494:pharmacology; D014746:Vimentin / Q000378:metabolism
D000952:Antigens, Polyomavirus Transforming; D003470:Culture Media; D014746:Vimentin; D007633:Keratins; D015291:Transforming Growth Factors; D002118:Calcium