Hepatic stimulator substance (HSS), also known as augmenter of liver regeneration (ALR), acts as a hepatotrophic growth factor to promote liver regeneration after liver damage or partial hepatectomy. However, the expression and function of HSS during liver development in mammals remain largely unknown. In this work, the hepatoblasts were isolated from mice at embryonic day 13.5 (E13.5), and HSS expression and its role during hepatoblast maturation were investigated. The results showed that HSS expression was enhanced in the hepatoblasts compared with mouse primary hepatocytes. HSS expression (23 kDa) was significantly decreased if the hepatoblast maturation was induced by a combination of oncostatin M (OSM), dexamethasone (DEX), and hepatocyte growth factor (HGF). We also found that knockdown of HSS expression (mainly 23-kDa isoform) by siRNA promoted hepatoblast maturation and also activated the signal transducer and activator of transcription 3 (STAT3) phosphorylation levels. However, if STAT3 activity was blocked by a small-molecule inhibitor Stattic, then hepatocyte maturation could be abolished, suggesting that STAT3 was most likely a potential molecule responsible for HSS signaling. In summary, our results demonstrated for the first time that HSS might be an active factor participating in the regulation of liver development and hepatocyte maturation.
Get full access to this article
View all access options for this article.
References
1.
JungJ. (1999). Initiation of mammalian liver development from endoderm by fibroblast growth factors. Science, 284:1998–2003.
2.
WattAJ, GarrisonWD and DuncanSA. (2003). HNF4: a central regulator of hepatocyte differentiation and function. Hepatology, 37:1249–1253.
3.
RastegarM, RousseauGG and LemaigreFP. (2000). CCAAT/enhancer-binding protein-alpha is a component of the growth hormone-regulated network of liver transcription factors. Endocrinology, 141:1686–1692.
4.
WestmacottA, BurkeZD, OliverG, SlackJM and ToshD. (2006). C/EBPalpha and C/EBPbeta are markers of early liver development. Int J Dev Biol, 50:653–657.
5.
GuoD, DongLY, WuY, YangL and AnW. (2008). Down-regulation of hepatic nuclear factor 4alpha on expression of human hepatic stimulator substance via its action on the proximal promoter in HepG2 cells. Biochem J, 415:111–121.
6.
DongLY, SunG, JiangL, ShaoL, HuY, JiangY, WangY and AnW. (2010). Epidermal growth factor down-regulates the expression of human hepatic stimulator substance via CCAAT/enhancer-binding protein beta in HepG2 cells. Biochem J, 431:277–287.
7.
LaBrecqueDR and PeschLA. (1975). Preparation and partial characterization of hepatic regenerative stimulator substance (SS) from rat liver. J Physiol, 248:273–284.
8.
LaBrecqueDR, SteeleG, FogertyS, WilsonM and BartonJ. (1987). Purification and physical-chemical characterization of hepatic stimulator substance. Hepatology, 7:100–106.
9.
LaBrecqueDR. (1991). Hepatic stimulator substance. Discovery, characteristics and mechanism of action. Dig Dis Sci, 36:669–673.
10.
FleigWE and HossG. (1989). Partial purification of rat hepatic stimulator substance and characterization of its action on hepatoma cells and normal hepatocytes. Hepatology, 9:240–248.
11.
HagiyaM, FrancavillaA, PolimenoL, IharaI, SakaiH, SekiT, ShimonishiM, PorterKA and StarzlTE. (1995). Cloning and sequence analysis of the rat augmenter of liver regeneration (ALR) gene: expression of biologically active recombinant ALR and demonstration of tissue distribution. Proc Natl Acad Sci U S A, 92:3076
12.
GiordaR, HagiyaM, SekiT, ShimonishiM, SakaiH, MichaelsonJ, FrancavillaA, StarzlTE and TruccoM. (1996). Analysis of the structure and expression of the augmenter of liver regeneration (ALR) gene. Mol Med, 2:97–108.
13.
WangG, YangX, ZhangY, WangQ, ChenH, WeiH, XingG, XieL, HuZ, et al. (1999). Identification and characterization of receptor for mammalian hepatopoietin that is homologous to yeast ERV1. J Biol Chem, 274:11469–11472.
14.
LiY, LiM, XingG, HuZ, WangQ, DongC, WeiH, FanG, ChenJ, et al. (2000). Stimulation of the mitogen-activated protein kinase cascade and tyrosine phosphorylation of the epidermal growth factor receptor by hepatopoietin. J Biol Chem, 275:37443–37447.
15.
LangeH, LisowskyT, GerberJ, MuhlenhoffU, KispalG and LillR. (2001). An essential function of the mitochondrial sulfhydryl oxidase Erv1p/ALR in the maturation of cytosolic Fe/S proteins. EMBO Rep, 2:715–720.
16.
WuCK, DaileyTA, DaileyHA, WangBC and RoseJP. (2003). The crystal structure of augmenter of liver regeneration: a mammalian FAD-dependent sulfhydryl oxidase. Protein Sci, 12:1109–1118.
17.
ThirunavukkarasuC, WangLF, HarveySA, WatkinsSC, ChailletJR, PrelichJ, StarzlTE and GandhiCR. (2008). Augmenter of liver regeneration: an important intracellular survival factor for hepatocytes. J Hepatol, 48:578–588.
18.
GandhiCR. (2012). Augmenter of liver regeneration. Fibrogenesis Tissue Repair, 5:10
19.
LiY, FarooqM, ShengD, ChandramouliC, LanT, MahajanNK, KiniRM, HongY, LisowskyT and GeR. (2012). Augmenter of liver regeneration (alr) promotes liver outgrowth during zebrafish hepatogenesis. PLoS One, 7:e30835
20.
TaoT and PengJ. (2009). Liver development in zebrafish (Danio rerio). J Genet Genomics, 36:325–334.
21.
DayoubR, GroitlP, DobnerT, BosserhoffAK, SchlittHJ and WeissTS. (2010). Foxa2 (HNF-3beta) regulates expression of hepatotrophic factor ALR in liver cells. Biochem Biophys Res Commun, 395:465–470.
22.
DayoubR, VogelA, SchuettJ, LupkeM, SpiekerSM, KetternN, HildtE, MelterM and WeissTS. (2013). Nrf2 activates augmenter of liver regeneration (ALR) via antioxidant response element and links oxidative stress to liver regeneration. Mol Med, 19:237–244.
23.
DabevaMD, PetkovPM, SandhuJ, OrenR, LaconiE, HurstonE and ShafritzDA. (2000). Proliferation and differentiation of fetal liver epithelial progenitor cells after transplantation into adult rat liver. Am J Pathol, 156:2017–2031.
24.
RoglerLE. (1997). Selective bipotential differentiation of mouse embryonic hepatoblasts in vitro. Am J Pathol, 150:591–602.
25.
de LonguevilleF, AtienzarFA, MarcqL, DufraneS, EvrardS, WoutersL, LerouxF, BertholetV, GerinB, et al. (2003). Use of a low-density microarray for studying gene expression patterns induced by hepatotoxicants on primary cultures of rat hepatocytes. Toxicol Sci, 75:378–392.
26.
LivakKJ and SchmittgenTD. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25:402–408.
27.
TilghmanSM and BelayewA. (1982). Transcriptional control of the murine albumin/alpha-fetoprotein locus during development. Proc Natl Acad Sci U S A, 79:5254–5257.
28.
CaoY, FuYL, YuM, YuePB, GeCH, XuWX, ZhanYQ, LiCY, LiW, et al. (2009). Human augmenter of liver regeneration is important for hepatoma cell viability and resistance to radiation-induced oxidative stress. Free Radic Biol Med, 47:1057–1066.
29.
ZhangW, LiW, LiuB, WangP, LiW and ZhangH. (2012). Efficient generation of functional hepatocyte-like cells from human fetal hepatic progenitor cells in vitro. J Cell Physiol, 227:2051–2058.
30.
KamiyaA, KinoshitaT and MiyajimaA. (2001). Oncostatin M and hepatocyte growth factor induce hepatic maturation via distinct signaling pathways. FEBS Lett, 492:90–94.
31.
HeF, WuC, TuQ and XingG. (1993). Human hepatic stimulator substance: a product of gene expression of human fetal liver tissue. Hepatology, 17:225–229.
32.
IvanovaNB, DimosJT, SchanielC, HackneyJA, MooreKA and LemischkaIR. (2002). A stem cell molecular signature. Science, 298:601–604.
33.
Ramalho-SantosM, YoonS, MatsuzakiY, MulliganRC and Melton.DA (2002). “Stemness”.: transcriptional profiling of embryonic and adult stem cells. Science, 298:597–600.
34.
TengEC, ToddLR, RibarTJ, LentoW, DimascioL, MeansAR and SankarU. (2011). Gfer inhibits Jab1-mediated degradation of p27kip1 to restrict proliferation of hematopoietic stem cells. Mol Biol Cell, 22:1312–1320.
35.
ToddLR, DaminMN, GomathinayagamR, HornSR, MeansAR and SankarU. (2010). Growth factor erv1-like modulates Drp1 to preserve mitochondrial dynamics and function in mouse embryonic stem cells. Mol Biol Cell, 21:1225–1236.
36.
HiroseY, ItohT and MiyajimaA. (2009). Hedgehog signal activation coordinates proliferation and differentiation of fetal liver progenitor cells. Exp Cell Res, 315:2648–2657.
37.
diIorioPJ, MossJB, SbrognaJL, KarlstromRO and MossLG. (2002). Sonic hedgehog is required early in pancreatic islet development. Dev Biol, 244:75–84.
38.
RoyS, QiaoT, WolffC and InghamPW. (2001). Hedgehog signaling pathway is essential for pancreas specification in the zebrafish embryo. Curr Biol, 11:1358–1363.
39.
FukadaT, HibiM, YamanakaY, Takahashi-TezukaM, FujitaniY, YamaguchiT, NakajimaK and HiranoT. (1996). Two signals are necessary for cell proliferation induced by a cytokine receptor gp130: involvement of STAT3 in anti-apoptosis. Immunity, 5:449–460.
40.
MatsuiT, KinoshitaT, HiranoT, YokotaT and MiyajimaA. (2002). STAT3 down-regulates the expression of cyclin D during liver development. J Biol Chem, 277:36167–36173.
41.
YamashitaS, MiyagiC, Carmany-RampeyA, ShimizuT, FujiiR, SchierAF and HiranoT. (2002). Stat3 controls cell movements during Zebrafish gastrulation. Dev Cell, 2:363–375.
42.
YangY, PanX, LeiW, WangJ, ShiJ, LiF and SongJ. (2006). Regulation of transforming growth factor-beta 1-induced apoptosis and epithelial-to-mesenchymal transition by protein kinase A and signal transducers and activators of transcription 3. Cancer Res, 66:8617–8624.
43.
ChoiSS and DiehlAM. (2009). Epithelial-to-mesenchymal transitions in the liver. Hepatology, 50:2007–2013.
44.
DayoubR, WagnerH, BatailleF, StoltzingO, SprussT, BuechlerC, SchlittHJ and WeissTS. (2011). Liver regeneration associated protein (ALR) exhibits antimetastatic potential in hepatocellular carcinoma. Mol Med, 17:221–228.
45.
ZhangH, DongLY, SunG and AnW. (2014). Downregulation of hepatic stimulator substance during the early phase of liver regeneration inhibits E-cadherin expression in mice. Int J Biochem Cell Biol, 47:38–46.
46.
TianZJ and AnW. (2004). ERK1/2 contributes negative regulation to STAT3 activity in HSS-transfected HepG2 cells. Cell Res, 14:141–147.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
