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Lymphoid enhancer-binding factor-1 promotes stemness and poor differentiation of hepatocellular carcinoma by directly activating the NOTCH pathway

Abstract

The poorly differentiated hepatocellular carcinoma (HCC) cells are usually characterized by immature hepatic progenitor cell-like properties, such as enhanced self-renewal ability, resistance to chemotherapeutic drugs, and a loss of mature hepatocyte proteins. However, the molecular mechanisms governing this process still remain unclear. In this study, we found the lymphoid enhancer-binding factor-1 (LEF1), a transcriptional factor, was frequently overexpressed in HCCs, which was significantly associated with poor prognosis and tumor cell differentiation. Functional studies have found that LEF1 enhanced cell growth, foci formation, colony formation in soft agar, and tumor formation in nude mice. Different from its canonical roles in the WNT signaling pathway, we found that LEF1 could activate the critical members (e.g., NOTCH1 and NOTCH2) of the NOTCH signaling pathway through directly binding to their promoter regions. Further studies have found that LEF1 could enhance the self-renewal ability, drug resistance, dedifferentiation, and invasion of HCC cells. The oncogenic functions and the effects of LEF1 on cancer stemness could be effectively inhibited by NOTCH inhibitor. Further characterization of LEF1 may lead to the development of novel therapeutic strategies for HCC treatment.

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References

  1. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.

    Article  Google Scholar 

  2. Tong MJ, Chavalitdhamrong D, Lu DS, Raman SS, Gomes A, Duffy JP, et al. Survival in Asian Americans after treatments for hepatocellular carcinoma: a seven-year experience at UCLA. J Clin Gastroenterol. 2010;44:e63–70.

    Article  Google Scholar 

  3. Dhanasekaran R, Bandoh S, Roberts LR. Molecular pathogenesis of hepatocellular carcinoma and impact of therapeutic advances. F1000Res. 2016;5:879.

    Article  Google Scholar 

  4. Okamura RM, Sigvardsson M, Galceran J, Verbeek S, Clevers H, Grosschedl R. Redundant regulation of T cell differentiation and TCRalpha gene expression by the transcription factors LEF-1 and TCF-1. Immunity. 1998;8:11–20.

    Article  CAS  Google Scholar 

  5. Reya T, O’Riordan M, Okamura R, Devaney E, Willert K, Nusse R, et al. Wnt signaling regulates B lymphocyte proliferation through a LEF-1 dependent mechanism. Immunity. 2000;13:15–24.

    Article  CAS  Google Scholar 

  6. Wu W, Zhu H, Fu Y, Shen W, Miao K, Hong M, et al. High LEF1 expression predicts adverse prognosis in chronic lymphocytic leukemia and may be targeted by ethacrynic acid. Oncotarget. 2016;7:21631–43.

    PubMed  PubMed Central  Google Scholar 

  7. Petropoulos K, Arseni N, Schessl C, Stadler CR, Rawat VP, Deshpande AJ, et al. A novel role for Lef-1, a central transcription mediator of Wnt signaling, in leukemogenesis. J Exp Med. 2008;205:515–22.

    Article  CAS  Google Scholar 

  8. Staal FJ, Luis TC, Tiemessen MM. WNT signalling in the immune system: WNT is spreading its wings. Nat Rev Immunol. 2008;8:581–93.

    Article  CAS  Google Scholar 

  9. Clevers H. Wnt/beta-catenin signaling in development and disease. Cell. 2006;127:469–80.

    Article  CAS  Google Scholar 

  10. McGregor SM The role of Lef1 in T cell development and lymphomagenesis[J]. Dissertations & Theses - Gradworks, 2009.

  11. Visvader JE, Lindeman GJ. Cancer stem cells: current status and evolving complexities. Cell Stem Cell. 2012;10:717–28.

    Article  CAS  Google Scholar 

  12. Kopan R, Ilagan MX. The canonical Notch signaling pathway: unfolding the activation mechanism. Cell. 2009;137:216–33.

    Article  CAS  Google Scholar 

  13. Ungerback J, Elander N, Grunberg J, Sigvardsson M, Soderkvist P. The Notch-2 gene is regulated by Wnt signaling in cultured colorectal cancer cells. PLoS ONE 2011;6:e17957.

    Article  Google Scholar 

  14. Heitzler P, Bourouis M, Ruel L, Carteret C, Simpson P. Genes of the Enhancer of split and achaete-scute complexes are required for a regulatory loop between Notch and Delta during lateral signalling in Drosophila. Development. 1996;122:161–71.

    CAS  PubMed  Google Scholar 

  15. Leimeister C, Externbrink A, Klamt B, Gessler M. Hey genes: a novel subfamily of hairy- and Enhancer of split related genes specifically expressed during mouse embryogenesis. Mech Dev. 1999;85:173–7.

    Article  CAS  Google Scholar 

  16. Kabos P, Kabosova A, Neuman T. Blocking HES1 expression initiates GABAergic differentiation and induces the expression ofp21(CIP1/WAF1) in human neural stem cells. J Biol Chem. 2002;277:8763–6.

    Article  CAS  Google Scholar 

  17. Weng AP, Millholland JM, Yashiro-Ohtani Y, Arcangeli ML, Lau A, Wai C, et al. c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev. 2006;20:2096–109.

    Article  CAS  Google Scholar 

  18. Oswald F, Liptay S, Adler G, Schmid RM. NF-kappaB2 is a putative target gene of activated Notch-1 via RBP-Jkappa. Mol Cell Biol. 1998;18:2077–88.

    Article  CAS  Google Scholar 

  19. Nakahara F, Kitaura J, Uchida T, Nishida C, Togami K, Inoue D, et al. Hes1 promotes blast crisis in chronic myelogenous leukemia through MMP-9 upregulation in leukemic cells. Blood. 2014;123:3932–42.

    Article  CAS  Google Scholar 

  20. Lai EC. Notch signaling: control of cell communication and cell fate. Development. 2004;131:965–73.

    Article  CAS  Google Scholar 

  21. Kopan R. Notch: a membrane-bound transcription factor. J Cell Sci. 2002;115:1095–7.

    CAS  PubMed  Google Scholar 

  22. Kohn A, Rutkowski TP, Liu Z, Mirando AJ, Zuscik MJ, O’Keefe RJ, et al. Notch signaling controls chondrocyte hypertrophy via indirect regulation of Sox9. Bone Res. 2015;3:15021.

    Article  CAS  Google Scholar 

  23. Sekiya S, Suzuki A. Intrahepatic cholangiocarcinoma can arise from Notch-mediated conversion of hepatocytes. J Clin Invest. 2012;122:3914–8.

    Article  CAS  Google Scholar 

  24. Valenti L, Mendoza RM, Rametta R, Maggioni M, Kitajewski C, Shawber CJ, et al. Hepatic notch signaling correlates with insulin resistance and nonalcoholic fatty liver disease. Diabetes. 2013;62:4052–62.

    Article  CAS  Google Scholar 

  25. Si-Tayeb K, Lemaigre FP, Duncan SA. Organogenesis and development of the liver. Dev Cell. 2010;18:175–89.

    Article  CAS  Google Scholar 

  26. Mishra L, Banker T, Murray J, Byers S, Thenappan A, He AR, et al. Liver stem cells and hepatocellular carcinoma. Hepatology. 2009;49:318–29.

    Article  Google Scholar 

  27. Park HJ, Choi YJ, Kim JW, Chun HS, Im I, Yoon S, et al. Differences in the epigenetic regulation of cytochrome P450 genes between human embryonic stem cell-derived hepatocytes and primary hepatocytes. PLoS ONE 2015;10:e0132992.

    Article  Google Scholar 

  28. Liu M, Chen L, Ma NF, Chow RK, Li Y, Song Y, et al. CHD1L promotes lineage reversion of hepatocellular carcinoma through opening chromatin for key developmental transcription factors. Hepatology. 2016;63:1544–59.

    Article  CAS  Google Scholar 

  29. Cook JJ, Wildsmith KR, Gilberto DB, Holahan MA, Kinney GG, Mathers PD, et al. Acute gamma-secretase inhibition of nonhuman primate CNS shifts amyloid precursor protein (APP) metabolism from amyloid-beta production to alternative APP fragments without amyloid-beta rebound. J Neurosci. 2010;30:6743–50.

    Article  CAS  Google Scholar 

  30. Kim K, Lu Z, Hay ED. Direct evidence for a role of beta-catenin/LEF-1 signaling pathway in induction of EMT. Cell Biol Int. 2002;26:463–76.

    Article  CAS  Google Scholar 

  31. Reya T, Clevers H. Wnt signalling in stem cells and cancer. Nature. 2005;434:843–50.

    Article  CAS  Google Scholar 

  32. Zhang Y, Yu J, Shi C, Huang Y, Wang Y, Yang T, et al. Lef1 contributes to the differentiation of bulge stem cells by nuclear translocation and cross-talk with the Notch signaling pathway. Int J Med Sci. 2013;10:738–46.

    Article  CAS  Google Scholar 

  33. Waterman ML. Lymphoid enhancer factor/T cell factor expression in colorectal cancer. Cancer Metastasis Rev. 2004;23:41–52.

    Article  CAS  Google Scholar 

  34. Galceran J, Sustmann C, Hsu SC, Folberth S, Grosschedl R. LEF1-mediated regulation of Delta-like1 links Wnt and Notch signaling in somitogenesis. Genes Dev. 2004;18:2718–23.

    Article  CAS  Google Scholar 

  35. Collu GM, Hidalgo-Sastre A, Brennan K. Wnt-Notch signalling crosstalk in development and disease. Cell Mol Life Sci. 2014;71:3553–67.

    Article  CAS  Google Scholar 

  36. Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development. Science. 1999;284:770–6.

    Article  CAS  Google Scholar 

  37. Ranganathan P, Weaver KL, Capobianco AJ. Notch signalling in solid tumours: a little bit of everything but not all the time. Nat Rev Cancer. 2011;11:338–51.

    Article  CAS  Google Scholar 

  38. Ji J, Wang XW. Clinical implications of cancer stem cell biology in hepatocellular carcinoma. Semin Oncol. 2012;39:461–72.

    Article  CAS  Google Scholar 

  39. Ma NF, Hu L, Fung JM, Xie D, Zheng BJ, Chen L, et al. Isolation and characterization of a novel oncogene, amplified in liver cancer 1, within a commonly amplified region at 1q21 in hepatocellular carcinoma. Hepatology. 2008;47:503–10.

    Article  CAS  Google Scholar 

  40. Chen L, Hu L, Chan TH, Tsao GS, Xie D, Huo KK, et al. Chromodomain helicase/adenosine triphosphatase DNA binding protein 1-like (CHD1l) gene suppresses the nucleus-to-mitochondria translocation of nur77 to sustain hepatocellular carcinoma cell survival. Hepatology. 2009;50:122–9.

    Article  CAS  Google Scholar 

  41. Chen L, Chan TH, Yuan YF, Hu L, Huang J, Ma S, et al. CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients. J Clin Invest. 2010;120:1178–91.

    Article  CAS  Google Scholar 

  42. Chen L, Yuan YF, Li Y, Chan TH, Zheng BJ, Huang J, et al. Clinical significance of CHD1L in hepatocellular carcinoma and therapeutic potentials of virus-mediated CHD1L depletion. Gut. 2011;60:534–43.

    Article  CAS  Google Scholar 

  43. Ma S, Lee TK, Zheng BJ, Chan KW, Guan XY. CD133 + HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway. Oncogene. 2008;27:1749–58.

    Article  CAS  Google Scholar 

  44. Fu L, Qin YR, Xie D, Hu L, Kwong DL, Srivastava G, et al. Characterization of a novel tumor-suppressor gene PLC delta 1 at 3p22 in esophageal squamous cell carcinoma. Cancer Res. 2007;67:10720–6.

    Article  CAS  Google Scholar 

  45. Vasaikar SV, Straub P, Wang J, Zhang B. LinkedOmics: analyzing multi-omics data within and across 32 cancer types. Nucleic Acids Res. 2017;46:D956–D96.

    Article  Google Scholar 

  46. Chandrashekar DS, Bashel B, Balasubramanya SAH, Creighton CJ, Ponce-Rodriguez I, Chakravarthi B, et al. UALCAN: a portal for facilitating tumor subgroup gene expression and survival analyses. Neoplasia. 2017;19:649–58.

    Article  CAS  Google Scholar 

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Funding

This work was supported by grants from the Hong Kong Research Grant Council (RGC), including GRF (17143716 and 767313), Collaborative Research Funds (C7038-14G and C7027-14G), Theme-based Research Scheme (T12-704/16-R), and National Natural Science Foundation of China (81772554, 81272416, and 81372583). This work was also supported by the Shenzhen Peacock Team Project (KQTD 2015033117210153). Professor XY Guan is a Sophie YM Chan Professor in Cancer Research.

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S.F. and X.Y.G. initiated and designed the study. S.F. wrote the manuscript with inputs from X.Y.G. and S.F. and M.L. designed the experiments and interpreted the results. S.F. performed all the experiments with assistance from M.L., L.L., F.F.Z., Y.L., Q.Y. and Y.Z.C., HCC clinical samples and the relevant clinical information were provided by Y.H.Z. and Y.F.Y. and X.Y.G. supervised the project.

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Correspondence to Xin-Yuan Guan.

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Fang, S., Liu, M., Li, L. et al. Lymphoid enhancer-binding factor-1 promotes stemness and poor differentiation of hepatocellular carcinoma by directly activating the NOTCH pathway. Oncogene 38, 4061–4074 (2019). https://doi.org/10.1038/s41388-019-0704-y

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