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The STAT3 inhibitor S3I-201 suppresses fibrogenesis and angiogenesis in liver fibrosis

Laboratory Investigationvolume 98pages16001613 (2018) | Download Citation


Liver fibrosis is a common pathological response to chronic hepatic injury. STAT3 is actively involved in the fibrogenesis and angiogenesis seen in liver fibrosis. S3I-201 (NSC 74859) is a chemical inhibitor of STAT3 activity, which blocks the dimerization of STAT3, STAT3-DNA binding and transcription activity. This study evaluated the effects of S3I-201 against liver fibrosis. S3I-201 inhibited the proliferation, migration, and actin filament formation in primary human hepatic stellate cells (HSCs), as well as the expression of α-SMA, collagen I and TIMP1 in both primary HSC and in a CCl4-induced fibrosis mouse model. S3I-201 induced both apoptosis and cell cycle arrest in the HSC cell line (LX-2). S3I-201 inhibited the expression of fibrogenesis factors TGFβ1 and TGFβRII, as well as the downstream phosphorylation of Smad2, Smad3, Akt and ERK induced by TGFβ1. In addition to fibrogenesis, both in vitro and in vivo assays showed that S3I-201 inhibited angiogenesis through expression suppression of VEGF and VEGFR2. Moreover, S3I-201 also had a synergistic effect with sorafenib, an FDA approved liver cancer drug, in the proliferation, apoptosis, angiogenesis and fibrogenesis of HSC. S3I-201 suppressed liver fibrosis through multiple mechanisms, and combined with sorafenib, S3I-201 could be a potentially effective antifibrotic agent.

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  1. 1.

    Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005;115:209–18.

  2. 2.

    Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev. 2017;121:27–42.

  3. 3.

    Trautwein C, Friedman SL, Schuppan D, et al. Hepatic fibrosis: concept to treatment. J Hepatol. 2015;62:15–24.

  4. 4.

    Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story. Gut. 2015;64:830–41.

  5. 5.

    Ellis EL, Mann DA. Clinical evidence for the regression of liver fibrosis (vol 56, pg 1171, 2012). J Hepatol. 2014;60:468–9.

  6. 6.

    Wasmuth HE, Tacke F, Trautwein C. Chemokines in liver inflammation and fibrosis. Semin Liver Dis. 2010;30:215–25.

  7. 7.

    Gressner AM, Weiskirchen R, Breitkopf K, et al. Roles of TGF-beta in hepatic fibrosis. Front Biosci. 2002;7:793–807.

  8. 8.

    Dooley S, Hamzavi J, Breitkopf K, et al. Smad7 prevents activation of hepatic stellate cells and liver fibrosis in rats. Gastroenterology. 2003;125:178–91.

  9. 9.

    Tang LY, Heller M, Meng Z, et al. Transforming growth factor-beta (TGF-beta) directly activates the JAK1-STAT3 axis to induce hepatic fibrosis in coordination with the SMAD pathway. J Biol Chem. 2017;292:4302–12.

  10. 10.

    Liu XJ, Hu H, Yin JQ. Therapeutic strategies against TGF-beta signaling pathway in hepatic fibrosis. Liver Int. 2006;26:8–22.

  11. 11.

    Xu MY, Hu JJ, Shen J, et al. Stat3 signaling activation crosslinking of TGF-beta 1 in hepatic stellate cell exacerbates liver injury and fibrosis. Bba-Mol Basis Dis. 2014;1842:2237–45.

  12. 12.

    Ogata H, Chinen T, Yoshida T, et al. Loss of SOCS3 in the liver promotes fibrosis by enhancing STAT3-mediated TGF-beta 1 production. Oncogene. 2006;25:2520–30.

  13. 13.

    Corpechot C, Barbu V, Wendum D, et al. Hypoxia-induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis. Hepatology. 2002;35:1010–21.

  14. 14.

    Yoshiji H, Kuriyama S, Yoshii J, et al. Vascular endothelial growth factor and receptor interaction is a prerequisite for murine hepatic fibrogenesis. Gut. 2003;52:1347–54.

  15. 15.

    Alvarez JV, Febbo PG, Ramaswamy S, et al. Identification of a genetic signature of activated signal transducer and activator of transcription 3 in human tumors. Cancer Res. 2005;65:5054–62.

  16. 16.

    Siddiquee K, Zhang S, Guida WC, et al. Selective chemical probe inhibitor of Stat3, identified through structure-based virtual screening, induces antitumor activity. Proc Natl Acad Sci USA. 2007;104:7391–6.

  17. 17.

    Liu YQ, Wang Z, Wang JN, et al. A histone deacetylase inhibitor, largazole, decreases liver fibrosis and angiogenesis by inhibiting transforming growth factor-beta and vascular endothelial growth factor signalling. Liver Int. 2013;33:504–15.

  18. 18.

    Liu YQ, Wang Z, Kwong SQ, et al. Inhibition of PDGF, TGF-beta, and Abl signaling and reduction of liver fibrosis by the small molecule Bcr-Abl tyrosine kinase antagonist Nilotinib. J Hepatol. 2011;55:612–25.

  19. 19.

    Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70:440–6.

  20. 20.

    Lin L, Amin R, Gallicano GI, et al. The STAT3 inhibitor NSC 74859 is effective in hepatocellular cancers with disrupted TGF-beta signaling. Oncogene. 2009;28:961–72.

  21. 21.

    Horiguchi N, Lafdil F, Miller AM, et al. Dissociation between liver inflammation and hepatocellular damage induced by carbon tetrachloride in myeloid cell-specific signal transducer and activator of Transcription 3 gene knockout mice. Hepatology. 2010;51:1724–34.

  22. 22.

    Deng YR, Ma HD, Tsuneyama K, et al. STAT3-mediated attenuation of CCl4-induced mouse liver fibrosis by the protein kinase inhibitor sorafenib. J Autoimmun. 2013;46:25–34.

  23. 23.

    Mair M, Zollner G, Schneller D, et al. Signal transducer and activator of transcription 3 protects from liver injury and fibrosis in a mouse model of sclerosing cholangitis. Gastroenterology. 2010;138:2499–508.

  24. 24.

    Plum W, Tschaharganeh DF, Kroy DC, et al. Lack of glycoprotein 130/signal transducer and activator of transcription 3-mediated signaling in hepatocytes enhances chronic liver injury and fibrosis progression in a model of sclerosing cholangitis. Am J Pathol. 2010;176:2236–46.

  25. 25.

    Wang H, Lafdil F, Kong XN, et al. Signal transducer and activator of transcription 3 in liver diseases: a novel therapeutic target. Int J Biol Sci. 2011;7:536–50.

  26. 26.

    Jiang JX, Mikami K, Venugopal S, et al. Apoptotic body engulfment by hepatic stellate cells promotes their survival by the JAK/STAT and Akt/NF-kappa B-dependent pathways. J Hepatol. 2009;51:139–48.

  27. 27.

    Grutter MG. Caspases: key players in programmed cell death. Curr Opin Struct Biol. 2000;10:649–55.

  28. 28.

    Novo E, Marra F, Zamara E, et al. Overexpression of Bcl-2 by activated human hepatic stellate cells: resistance to apoptosis as a mechanism of progressive hepatic fibrogenesis in humans. Gut. 2006;55:1174–82.

  29. 29.

    Fabregat I, Moreno-Caceres J, Sanchez A, et al. TGF-beta signalling and liver disease. Febs J. 2016;283:2219–32.

  30. 30.

    Lechuga CG, Hernandez-Nazara ZH, Rosales JAD, et al. TGF-beta 1 modulates matrix metalloproteinase-13 expression in hepatic stellate cells by complex mechanisms involving p38MAPK, PI3-kinase, AKT, andp70(S6k). Am J Physiol Gastrointest Liver Physiol. 2004;287:974–87.

  31. 31.

    Reif S, Lang A, Lindquist JN, et al. The role of focal adhesion kinase-phosphatidylinositol 3-kinase-Akt signaling in hepatic stellate cell proliferation and type I collagen expression. J Biol Chem. 2003;278:8083–90.

  32. 32.

    Feng XH, Derynck R. Specificity and versatility in TGF-beta signaling through Smads. Annu Rev Cell Dev Bi. 2005;21:659–93.

  33. 33.

    Wang G, Yu Y, Sun C, et al. STAT3 selectively interacts with Smad3 to antagonize TGF-beta signalling. Oncogene. 2016;35:4388–98.

  34. 34.

    Lee JS, Semela D, Iredale J, et al. Sinusoidal remodeling and angiogenesis: a new function for the liver-specific pericyte? Hepatology. 2007;45:817–25.

  35. 35.

    Chen XH, Yang GX, Song JH, et al. Probiotic yeast inhibits VEGFR signaling and angiogenesis in intestinal inflammation. PLoS ONE. 2013;8:e64227.

  36. 36.

    Yang L, Kwon J, Popov Y, et al. Vascular endothelial growth factor promotes fibrosis resolution and repair in mice. Gastroenterology. 2014;146:1339–50.

  37. 37.

    Nakamura I, Zakharia K, Banini BA, et al. Brivanib attenuates hepatic fibrosis in vivo and stellate cell activation in vitro by inhibition of FGF, VEGF and PDGF Signaling. PLoS ONE. 2015;10:e0142355.

  38. 38.

    Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. Nat Med. 2003;9:669–76.

  39. 39.

    Yang SY, Xin XH, Zlot C, et al. Vascular endothelial cell growth factor-driven endothelial tube formation is mediated by vascular endothelial cell growth factor receptor-2, a kinase insert domain-containing receptor. Arter Throm Vas. 2001;21:1934–40.

  40. 40.

    Varela-Rey M, Montiel-Duarte C, Oses-Prieto JA, et al. p38 MAPK mediates the regulation of alpha 1(I) procollagen mRNA levels by TNF-alpha and TGF-beta in a cell line of rat hepatic stellate cells. FEBS Lett. 2002;528:133–8.

  41. 41.

    Liu YQ, Wen XM, Lui ELH, et al. Therapeutic targeting of the PDGF and TGF-beta-signaling pathways in hepatic stellate cells by PTK787/ZK22258. Lab Invest. 2009;89:1152–60.

  42. 42.

    Gurbuz V, Konac E, Varol N, et al. Effects of AG490 and S3I-201 on regulation of the JAK/STAT3 signaling pathway in relation to angiogenesis in TRAIL-resistant prostate cancer cells in vitro. Oncol Lett. 2014;7:755–63.

  43. 43.

    Pang MY, Ma L, Gong RJ, et al. A novel STAT3 inhibitor, S3I-201, attenuates renal interstitial fibroblast activation and interstitial fibrosis in obstructive nephropathy. Kidney Int. 2010;78:257–68.

  44. 44.

    Wilhelm S, Carter C, Lynch M, et al. Discovery and development of sorafenib: a multikinase inhibitor for treating cancer. Nat Rev Drug Discov. 2006;5:835–44.

  45. 45.

    Wang Y, Gao JC, Zhang D, et al. New insights into the antifibrotic effects of sorafenib on hepatic stellate cells and liver fibrosis. J Hepatol. 2010;53:132–44.

  46. 46.

    Su TH, Shiau CW, Jao P, et al. Sorafenib and its derivative SC-1 exhibit antifibrotic effects through signal transducer and activator of transcription 3 inhibition. Proc Natl Acad Sci USA. 2015;112:7243–8.

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This work was partly supported by Guangdong Science and Technology Program (2017B030301018), research grants from Shenzhen Science and Technology Innovation Committee (JCYJ20160608140912962 and ZDSYS20140509142721429) and National Natural Science Foundation of China (31670753).

Author information


  1. College of Life Sciences, Nankai University, Tianjin, 300071, China

    • Zhuo Wang
    •  & Jiafu Long
  2. Department of Biology, Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Shenzhen Key Laboratory of Cell Microenvironment and SUSTech-HKU Joint Laboratories for Matrix Biology and Diseases, Southern University of Science and Technology, Shenzhen, 518055, China

    • Zhuo Wang
    • , Jia’an Li
    • , Wen’ang Xiao
    •  & Hongmin Zhang
  3. State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China

    • Jiafu Long


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The authors declare no conflict of interest.

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Correspondence to Zhuo Wang or Hongmin Zhang.

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