Familial combined hyperlipidemia plasma stimulates protein secretion by HepG2 cells: identification of fibronectin in the differential secretion proteome

J Lipid Res. 2002 Nov;43(11):1846-54. doi: 10.1194/jlr.m100441-jlr200.

Abstract

The aim of this study was to evaluate whether soluble factors in plasma of familial combined hyperlipidemia (FCHL) patients affect hepatic protein secretion. Cultured human hepatocytes, i.e., HepG2 cells, were incubated with fasting plasma (20%, v/v, in DMEM) from untreated FCHL patients or normolipidemic controls. Overall protein secretion was 10-15% higher after incubation with FCHL plasma. This was specifically caused by an increase in four secreted proteins, with estimated sizes of 240, 180, 120, and <40 kD (P < 0.001, P < 0.006, P < 0.002, P < 0.02, respectively). The 240 kD protein in the secretion proteome was identified as fibronectin by mass spectrometry. Plasma fibronectin concentrations were elevated in FCHL patients, confirming biological relevance of these data. Overall protein secretion by HepG2 cells correlated with concentrations of triglycerides (r = 0.61, P < 0.001) in the applied plasma samples. VLDL+IDL isolated from FCHL patients, induced a higher protein secretion than lipoproteins isolated from controls (P < 0.001). Remarkably, secretion of apoB, the structural protein of VLDL, was stimulated to a similar extent by FCHL and control plasma. FCHL plasma did not induce excess secretion of apoB by HepG2 cells compared with control plasma. FCHL plasma did stimulate secretion of several distinct hepatic proteins, among which fibronectin was identified.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apolipoproteins B / blood
  • Cholesterol / blood
  • Fatty Acids / blood
  • Fibronectins / blood
  • Fibronectins / chemistry
  • Fibronectins / metabolism*
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism*
  • Humans
  • Hyperlipidemia, Familial Combined / blood*
  • Insulin / blood
  • Lipoproteins, VLDL / pharmacology
  • Liver / pathology
  • Molecular Weight
  • Proteome / chemistry
  • Proteome / metabolism*
  • Triglycerides / blood
  • Tumor Cells, Cultured

Substances

  • Apolipoproteins B
  • Fatty Acids
  • Fibronectins
  • Insulin
  • Lipoproteins, VLDL
  • Proteome
  • Triglycerides
  • Cholesterol