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(Arteriosclerosis, Thrombosis, and Vascular Biology. 1997;17:2395-2404.)
© 1997 American Heart Association, Inc.

Articles

Quantitation of Platelet-Derived Growth Factor Receptors in Human Arterial Smooth Muscle Cells In Vitro

Alexandra Krettek; Gunnar Fager; Peter Jernberg; Gunnel Östergren-Lundén; ; Florentyna Lustig

From The Wallenberg Laboratory for Cardiovascular Research, Göteborg University, Sweden.

Correspondence to Alexandra Krettek, The Wallenberg Laboratory for Cardiovascular Research, Göteborg University, Sahlgrenska University Hospital, S-413 45 Göteborg, Sweden. E-mail alexandra.krettek{at}wlab.wall.gu.se

Abstract Platelet-derived growth factor (PDGF) is suggested to play an important role in the development of atherosclerosis as a migratory and mitogenic stimulus to arterial smooth muscle cells (ASMCs). Stimulated and unstimulated ASMCs were studied with respect to PDGF receptor (PDGF-R) mRNA and protein expression. Quantitative RT-PCR was developed for simultaneous evaluation of both PDGF-R and -Rß mRNA expression and a quantitative ELISA for estimation of corresponding PDGF-R subunits. On the mRNA level, the overall PDGF-Rß expression was approximately 100 times lower than that of PDGF-R. Furthermore, although PDGF-R mRNA levels were high irrespective of hASMC phenotype, PDGF-Rß mRNA was influenced by serum stimulation with lower copy numbers in proliferating and confluent cells compared with quiescent cells. On the protein level, quiescent hASMCs expressed 10 times more PDGF-Rß than PDGF-R. Serum stimulation decreased cell surface PDGF-Rs, with most prominent loss of PDGF-R (ELISA and immunohistochemistry). Our results suggest a differential regulatory pattern for PDGF-R and -Rß and are compatible with the usage of alternative promoters for regulation of -R expression. Further, it seems that the number of available receptor subunits is not the only determinant of variations in cell stimulation with different PDGF isoforms.

Key Words: PDGF receptor • cell proliferation • smooth muscle cell • PDGF

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