Establishment of human fibroma cell lines from a MEN1 patient by introduction of either hTERT or SV40 early region.
Establishment of tumor cell lines as model systems for studying tumor biology or as a part of immunotherapeutic anti-cancer strategies is of high importance, whereby the highest possible preservation of the original tumor cell phenotype is a prerequisite for these aims. Since overexpression of the catalytic subunit of human telomerase (hTERT) is known to minimally alter the cellular phenotype, we focused on the establishment of cell lines derived from human fibroma from a MEN1 patient by ectopic expression of hTERT. Additionally, a cell line was generated by introduction of the early region of SV40 (SV40 ER). Both approaches resulted in continuous cell lines, and neither T1-LOHG (hTERT) nor SV1-LOHG (SV40 ER) showed a transformed phenotype. While SV40 ER-transfected cells underwent dramatic changes in morphology and growth characteristics, hTERT-expressing cells indeed retained a phenotype highly similar to the parental cells. Nevertheless, hTERT overexpression resulted in increased growth rates after about 70 population doublings (PD) and alterations of mRNA levels of genes associated with tumor pathogenesis. Thus, our data suggest that ectopic hTERT expression leads to immortalization of LOHG-F, sustaining many characteristics of the non-transfected counterparts, but continuous growth in vitro is associated with changes of the cellular phenotype.
Voglauer
R
R
Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, 1190 Vienna, Austria. r.voglauer@iam.boku.ac.at
Grillari
J
J
Fortschegger
K
K
Wieser
M
M
Sterovsky
T
T
Gunsberg
P
P
Katinger
H
H
Pfragner
R
R
D000952:Antigens, Polyomavirus Transforming / Q000276:immunology*; D049109:Cell Proliferation; D002470:Cell Survival; D004268:DNA-Binding Proteins; D005350:Fibroma / Q000235:genetics* / Q000473:pathology*; D006801:Humans; D018761:Multiple Endocrine Neoplasia Type 1 / Q000235:genetics* / Q000473:pathology*; D010641:Phenotype; D012333:RNA, Messenger / Q000096:biosynthesis; D019098:Telomerase / Q000096:biosynthesis* / Q000494:pharmacology*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000952:Antigens, Polyomavirus Transforming; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D019098:Telomerase