Effect of platelet-derived and endothelial cell-derived growth factors on the low density lipoprotein receptor pathway in cultured human fibroblasts.

Human platelet-derived growth factor (PDGF) has been previously shown to stimulate low density lipoprotein (LDL) receptor activity in cultured cells. Studies were conducted to delineate in detail the effects of PDGF on the LDL receptor pathway in normal human fibroblasts and to explore relationships between the effects of PDGF on LDL metabolism, on cholesterol metabolism, and on DNA synthesis. Increasing concentrations of PDGF stimulated parallel increases in both DNA synthesis and 125I-LDL cell surface binding. The effect of PDGF was due entirely to an increase (up to 4.3-fold) in the number of receptor sites per cell, and not to a change in receptor affinity (Kd approximately 2.0 nM). Parallel PDGF concentration-dependent increases in 125I-LDL binding, internalization, and degradation at 37 degrees C were observed. The results indicate that PDGF-stimulated cells metabolize receptor-bound LDL in a manner that is identical with that seen with quiescent cells. A single study with highly purified PDGF demonstrated that it was PDGF itself, and not some other component in the partially purified PDGF preparation used in most of this work, that was responsible for the observed effects. Studies were conducted on the effects of PDGF on hydroxymethylglutaryl CoA reductase activity, on cholesterol esterification, and on down-regulation by LDL of the LDL receptor. These studies indicated that LDL cholesterol taken into the PDGF-stimulated cell via the receptor pathway, appeared to become available normally and to have metabolic effects within the cell similar to those seen in quiescent cells. Fibroblasts from subjects with familial hypercholesterolemia showed a normal mitogenic response to PDGF, despite the absence or near absence of an effect on the LDL receptor pathway. Finally, studies were also conducted with endothelial cell-conditioned medium (ECCM), used as a source of the endothelial cell-derived growth factor. ECCM was similar to PDGF in stimulating LDL binding, but differed strikingly from PDGF in that the degradation of internalized LDL was inhibited. As a result, ECCM-treated cells did not effectively increase cholesterol esterification or suppress hydroxymethylglutaryl CoA reductase activity when LDL was present. These findings with substances produced by endothelial cells may have important implications for atherogenesis.