A two‐dimensional electrophoresis preliminary approach to human hepatocarcinoma differentiation induced by PPAR‐agonists

• Published in: Journal of cellular and molecular medicine Vol. 9; no. 2; pp. 462 - 467
• Main Authors: Bottoni, Patrizia, Giardina, B., Martorana, G. E., Zuppi, Cecilia, Sole, P. De, Rossi, Cristina, Scatena, R.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.04.2005; John Wiley & Sons, Inc
• Subjects: Agonists; Albumin; alpha-Fetoproteins - metabolism; Bezafibrate - pharmacology; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Catalase - metabolism; Cell culture; Cell differentiation; Cell Differentiation - drug effects; Cell Line, Tumor; Cell Proliferation - drug effects; Clofibric Acid - pharmacology; Electron transport chain; Electrophoresis, Gel, Two-Dimensional; Enzymatic activity; Gemfibrozil - pharmacology; Heat shock proteins; Hepatocarcinoma differentiation; Hepatocellular carcinoma; HSP70 Heat-Shock Proteins - metabolism; Hsp70 protein; Humans; Hypoglycemic Agents - pharmacology; Hypolipidemic Agents - pharmacology; Lactic Acid - metabolism; Liver cancer; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Magnetic resonance spectroscopy; Metabolism; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; NADH Dehydrogenase - metabolism; Oxidative metabolism; Peroxisome proliferator-activated receptors; Peroxisome Proliferator-Activated Receptors - agonists; Phenotypes; Proteomics; Proteomics - methods; Respiration; Serum Albumin - metabolism; Thiazolidinediones - pharmacology; Transferrin - metabolism; Transferrins; Tumor markers; α-Fetoprotein
• ISSN: 1582-1838; 1582-4934
• DOI: 10.1111/j.1582-4934.2005.tb00371.x

Adopting biochemical and proteomic approaches, we investigated the effect of some PPAR‐agonists, a new class of differentiating agents, on human hepatocellular carcinoma Hep‐G2 cell line. Cancer differentiation was assayed by checking albumin, transferrin and α‐fetoprotein synthesis. Cell metabolism was studied by NMR spectroscopy of cell culture supernatants and by evaluation of mitochondrial respiratory chain enzyme activities. The two dimensional electrophoresis approach was employed to analyze modifications in the expression of cellular proteins linked to cell phenotype differentiation in the attempt to identify potential diagnostic and prognostic biomarkers of hepatocellular carcinoma. Results indicate that PPAR‐agonists are able to act as differentiating inducers in human hepatocellular carcinoma Hep‐G2 cell line as well as to inhibit mitochondrial respiratory chain Complex I, provoking a selective derangement of cellular oxidative metabolism. Lastly, two dimensional electrophoresis showed interesting modifications in the pattern of expression of cellular proteins that confirm biochemical data (increase in albumin and transferrin, decrease of alpha ‐fetoprotein synthesis) and, moreover, emphasize the meaning of these data by the increase of spots indicatively ascribed to HSP70 and catalase.