title: Suppressor of Cytokine Signaling-3 (SOCS-3) Induces Proprotein Convertase Subtilisin Kexin Type 9 (PCSK9) Expression in Hepatic HepG2 Cell Line.
abstract: The suppressor of cytokine signaling (SOCS) proteins are negative regulators of the JAK/STAT pathway activated by proinflammatory cytokines, including the tumor necrosis factor-α (TNF-α). SOCS3 is also implicated in hypertriglyceridemia associated to insulin resistance. Proprotein convertase subtilisin kexin type 9 (PCSK9) levels are frequently found to be positively correlated to insulin resistance and plasma very low density lipoprotein (VLDL) triglycerides concentrations. The present study aimed to investigate the possible role of TNF-α and JAK/STAT pathway on de novo lipogenesis and PCSK9 expression in HepG2 cells. TNF-α induced both SOCS3 and PCSK9 in a concentration-dependent manner. This effect was inhibited by transfection with siRNA anti-STAT3, suggesting the involvement of the JAK/STAT pathway. Retroviral overexpression of SOCS3 in HepG2 cells (HepG2(SOCS3)) strongly inhibited STAT3 phosphorylation and induced PCSK9 mRNA and protein, with no effect on its promoter activity and mRNA stability. Consistently, siRNA anti-SOCS3 reduced PCSK9 mRNA levels, whereas an opposite effect was observed with siRNA anti-STAT3. In addition, HepG2(SOCS3) express higher mRNA levels of key enzymes involved in the de novo lipogenesis, such as fatty-acid synthase, stearoyl-CoA desaturase (SCD)-1, and apoB. These responses were associated with a significant increase of SCD-1 protein, activation of sterol regulatory element-binding protein-1c (SREBP-1), accumulation of cellular triglycerides, and secretion of apoB. HepG2(SOCS3) show lower phosphorylation levels of insulin receptor substrate 1 (IRS-1) Tyr(896) and Akt Ser(473) in response to insulin. Finally, insulin stimulation produced an additive effect with SOCS3 overexpression, further inducing PCSK9, SREBP-1, fatty acid synthase, and apoB mRNA. In conclusion, our data candidate PCSK9 as a gene involved in lipid metabolism regulated by proinflammatory cytokine TNF-α in a SOCS3-dependent manner.
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Ruscica
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Ricci
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Macchi
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy, Centro per lo Studio delle Malattie Dismetaboliche e delle Iperlipemie-Enrica Grossi Paoletti, Università degli Studi di Milano, 20162 Milan, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Magni
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy, Centro di Eccellenza per le Malattie Neurodegenerative, Università degli Studi di Milano, Milan, Italy, and.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cristofani
authors.affiliation: Department of Veterans Affairs, Palo Alto Health Care System, 94304 Palo Alto, California.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Liu
authors.affiliation: From the Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, 20133 Milan, Italy, Multimedica IRCCS, 20099 Milan, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Corsini
authors.affiliation: Dipartimento di Scienze del Farmaco, Università degli Studi di Padova, 35131 Padua, Italy nicola.ferri@unipd.it.
authors.forename: Nicola
authors.initials: N
authors.lastname: Ferri
mesh_terms: D000818:Animals; D004790:Enzyme Induction; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007333:Insulin Resistance; D053612:Janus Kinases / Q000737:chemistry / Q000378:metabolism; D050155:Lipogenesis; D008297:Male; D008810:Mice, Inbred C57BL; D009154:Mutation; D009765:Obesity / Q000201:enzymology / Q000378:metabolism; D010766:Phosphorylation; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C106644:SOCS3 protein, human; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D053612:Janus Kinases; C472125:PCSK9 protein, human; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D012697:Serine Endopeptidases
keywords: HepG2; hepatocyte; insulin; proprotein convertase subtilisin/kexin type 9 (PCSK9); stat-3; suppressor of cytokine signaling 3 (SOCS3); tumor necrosis factor (TNF)