Rat vascular smooth muscle cells immortalized with SV40 large T antigen possess defined smooth muscle cell characteristics including growth inhibition by heparin.
Rat aortic smooth muscle cells (SMC) have been established by retroviral delivery of the complementary DNA (cDNA) for the simian virus 40 large T antigen (SV40LT) and examined for SMC phenotypic markers and growth characteristics, including responsiveness to the antiproliferative effects of heparin. The transfected cells (SV40LT-SMC) maintain defined SMC characteristics for more than 215 population doublings (PD) as judged by muscle-specific actin expression and growth inhibition by heparin. SV40LT-SMC greater than 129 PD become transformed while SV40LT-SMC less than 77 PD resemble nontransfected SMC morphologically and are nontumorigenic. SV40LT-SMC apparently release a growth factor which acts in an autocrine fashion, since (1) suramin inhibits SV40LT-SMC proliferation, (2) SV40LT-SMC-conditioned medium (CM) contains mitogenic activity, and (3) SV40LT-SMC CM suppresses the binding of platelet-derived growth factor to SMC. Heparin (10-100 micrograms/ml) is a potent inhibitor of both early (less than 80 PD) and late-passage (greater than 80 PD) SV40LT-SMC proliferation. The antiproliferative effects of heparin are similar to those previously observed for SMC by several criteria; the dose-response inhibition curves are indistinguishable from those obtained with nontransfected cells, other glycosaminoglycans have little effect on SV40LT-SMC growth, the antiproliferative effects of heparin are reversed in the presence of epidermal growth factor, and heparin displays high-affinity saturable binding to SV40LT-SMC. In conclusion, SV40LT-SMC are a continuous line of SMC-like cells that are sensitive to the growth inhibitor, heparin. SV40LT-SMC should facilitate studies of heparin inhibition and may be applicable for the study of other SMC characteristics as well.
Reilly
C F
CF
Department of Pharmacology, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
D000199:Actins / Q000378:metabolism; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001012:Aorta, Abdominal; D001769:Blood; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors; D006493:Heparin / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D011919:Rats, Inbred Strains; D011923:Rats, Nude; D012190:Retroviridae / Q000235:genetics; D013539:Simian virus 40 / Q000276:immunology*; D014162:Transfection
D000199:Actins; D000952:Antigens, Polyomavirus Transforming; D006493:Heparin; D004247:DNA