Uncovering and deciphering the pro-invasive role of HACE1 in melanoma cells

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HACE1 is an E3 ubiquitin ligase described as a tumour suppressor because HACE1-knockout mice develop multi-organ, late-onset cancers and because HACE1 expression is lost in several neoplasms, such as Wilms’ tumours and colorectal cancer. However, a search of public databases indicated that HACE1 expression is maintained in melanomas. We demonstrated that HACE1 promoted melanoma cell migration and adhesion in vitro and was required for mouse lung colonisation by melanoma cells in vivo. Transcriptomic analysis of HACE1-depleted melanoma cells revealed an inhibition of ITGAV and ITGB1 as well changes in other genes involved in cell migration. We revealed that HACE1 promoted the K27 ubiquitination of fibronectin and regulated its secretion. Secreted fibronectin regulated ITGAV and ITGB1 expression, as well as melanoma cell adhesion and migration. Our findings disclose a novel molecular cascade involved in the regulation of fibronectin secretion, integrin expression and melanoma cell adhesion. By controlling this cascade, HACE1 displays pro-tumoural properties and is an important regulator of melanoma cell invasive properties.

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  1. 1.

    Anglesio MS, Evdokimova V, Melnyk N, Zhang L, Fernandez CV, Grundy PE, et al. Differential expression of a novel ankyrin containing E3 ubiquitin-protein ligase, Hace1, in sporadic Wilms’ tumor versus normal kidney. Hum Mol Genet. 2004;13:2061–74.

  2. 2.

    Zhang L, Anglesio MS, O’Sullivan M, Zhang F, Yang G, Sarao R, et al. The E3 ligase HACE1 is a critical chromosome 6q21 tumor suppressor involved in multiple cancers. Nat Med. 2007;13:1060–9.

  3. 3.

    Torrino S, Visvikis O, Doye A, Boyer L, Stefani C, Munro P, et al. The E3 ubiquitin-ligase HACE1 catalyzes the ubiquitylation of active Rac1. Dev Cell. 2011;21:959–65.

  4. 4.

    Castillo-Lluva S, Tan CT, Daugaard M, Sorensen PH, Malliri A. The tumour suppressor HACE1 controls cell migration by regulating Rac1 degradation. Oncogene. 2013;32:1735–42.

  5. 5.

    Liu Z, Chen P, Gao H, Gu Y, Yang J, Peng H, et al. Ubiquitylation of autophagy receptor Optineurin by HACE1 activates selective autophagy for tumor suppression. Cancer Cell. 2014;26:106–20.

  6. 6.

    Hodis E, Watson IR, Kryukov GV, Arold ST, Imielinski M, Theurillat JP, et al. A landscape of driver mutations in melanoma. Cell. 2012;150:251–63.

  7. 7.

    Halaban R. RAC1 and melanoma. Clin Ther. 2015;37:682–5.

  8. 8.

    Mischiati C, Natali PG, Sereni A, Sibilio L, Giorda E, Cappellacci S, et al. cDNA-array profiling of melanomas and paired melanocyte cultures. J Cell Physiol. 2006;207:697–705.

  9. 9.

    Scatolini M, Grand MM, Grosso E, Venesio T, Pisacane A, Balsamo A, et al. Altered molecular pathways in melanocytic lesions. Int J Cancer. 2010;126:1869–81.

  10. 10.

    Goka ET, Lippman ME. Loss of the E3 ubiquitin ligase HACE1 results in enhanced Rac1 signaling contributing to breast cancer progression. Oncogene. 2015;34:5395–405.

  11. 11.

    Tortola L, Nitsch R, Bertrand MJ, Kogler M, Redouane Y, Kozieradzki I, et al. The tumor suppressor Hace1 is a critical regulator of TNFR1-mediated cell fate. Cell Rep. 2016;16:3414.

  12. 12.

    Caswell PT, Spence HJ, Parsons M, White DP, Clark K, Cheng KW, et al. Rab25 associates with alpha5beta1 integrin to promote invasive migration in 3D microenvironments. Dev Cell. 2007;13:496–510.

  13. 13.

    Morgan MR, Byron A, Humphries MJ, Bass MD. Giving off mixed signals--distinct functions of alpha5beta1 and alphavbeta3 integrins in regulating cell behaviour. IUBMB Life. 2009;61:731–8.

  14. 14.

    Digiacomo G, Tusa I, Bacci M, Cipolleschi MG, Dello Sbarba P, Rovida E. Fibronectin induces macrophage migration through a SFK-FAK/CSF-1R pathway. Cell Adh Migr. 2017;11:1–11.

  15. 15.

    Hibi K, Sakata M, Sakuraba K, Shirahata A, Goto T, Mizukami H, et al. Aberrant methylation of the HACE1 gene is frequently detected in advanced colorectal cancer. Anticancer Res. 2008;28:1581–4.

  16. 16.

    Oberoi-Khanuja TK, Rajalingam K. Ubiquitination of Rac1 by inhibitors of apoptosis (IAPs). Methods Mol Biol. 2014;1120:43–54.

  17. 17.

    Oberoi TK, Dogan T, Hocking JC, Scholz RP, Mooz J, Anderson CL, et al. IAPs regulate the plasticity of cell migration by directly targeting Rac1 for degradation. EMBO J. 2012;31:14–28.

  18. 18.

    Zhao J, Mialki RK, Wei J, Coon TA, Zou C, Chen BB, et al. SCF E3 ligase F-box protein complex SCF(FBXL19) regulates cell migration by mediating Rac1 ubiquitination and degradation. FASEB J. 2013;27:2611–9.

  19. 19.

    Tang D, Xiang Y, De Renzis S, Rink J, Zheng G, Zerial M, et al. The ubiquitin ligase HACE1 regulates Golgi membrane dynamics during the cell cycle. Nat Commun. 2011;2:501.

  20. 20.

    Sako N, Dessirier V, Bagot M, Bensussan A, Schmitt C. HACE1, a potential tumor suppressor gene on 6q21, is not involved in extranodal natural killer/T-cell lymphoma pathophysiology. Am J Pathol. 2014;184:2899–907.

  21. 21.

    Hollstein R, Parry DA, Nalbach L, Logan CV, Strom TM, Hartill VL, et al. HACE1 deficiency causes an autosomal recessive neurodevelopmental syndrome. J Med Genet. 2015;52:797–803.

  22. 22.

    van der Horst G, Bos L, van der Mark M, Cheung H, Heckmann B, Clement-Lacroix P, et al. Targeting of alpha-v integrins reduces malignancy of bladder carcinoma. PLoS ONE. 2014;9:e108464.

  23. 23.

    Carriero MV, Franco P, Votta G, Longanesi-Cattani I, Vento MT, Masucci MT, et al. Regulation of cell migration and invasion by specific modules of uPA: mechanistic insights and specific inhibitors. Curr Drug Targets. 2011;12:1761–71.

  24. 24.

    Elfenbein A, Simons M. Syndecan-4 signaling at a glance. J Cell Sci. 2013;126:3799–804.

  25. 25.

    Hapke S, Kessler H, Arroyo de Prada N, Benge A, Schmitt M, Lengyel E, et al. Integrin alpha(v)beta(3)/vitronectin interaction affects expression of the urokinase system in human ovarian cancer cells. J Biol Chem. 2001;276:26340–8.

  26. 26.

    Hofmann G, Bernabei PA, Crociani O, Cherubini A, Guasti L, Pillozzi S, et al. HERG K + channels activation during beta(1) integrin-mediated adhesion to fibronectin induces an up-regulation of alpha(v)beta(3) integrin in the preosteoclastic leukemia cell line FLG 29.1. J Biol Chem. 2001;276:4923–31.

  27. 27.

    Landowski TH, Olashaw NE, Agrawal D, Dalton WS. Cell adhesion-mediated drug resistance (CAM-DR) is associated with activation of NF-kappa B (RelB/p50) in myeloma cells. Oncogene. 2003;22:2417–21.

  28. 28.

    Milner R, Campbell IL. The extracellular matrix and cytokines regulate microglial integrin expression and activation. J Immunol. 2003;170:3850–8.

  29. 29.

    Brentnall M, Weir DB, Rongvaux A, Marcus AI, Boise LH. Procaspase-3 regulates fibronectin secretion and influences adhesion, migration and survival independently of catalytic function. J Cell Sci. 2014;127:2217–26.

  30. 30.

    Palicharla VR, Maddika S. HACE1 mediated K27 ubiquitin linkage leads to YB-1 protein secretion. Cell Signal. 2015;27:2355–62.

  31. 31.

    Jin YJ, Park I, Hong IK, Byun HJ, Choi J, Kim YM, et al. Fibronectin and vitronectin induce AP-1-mediated matrix metalloproteinase-9 expression through integrin alpha(5)beta(1)/alpha(v)beta(3)-dependent Akt, ERK and JNK signaling pathways in human umbilical vein endothelial cells. Cell Signal. 2011;23:125–34.

  32. 32.

    Matsuo M, Sakurai H, Ueno Y, Ohtani O, Saiki I. Activation of MEK/ERK and PI3K/Akt pathways by fibronectin requires integrin alphav-mediated ADAM activity in hepatocellular carcinoma: a novel functional target for gefitinib. Cancer Sci. 2006;97:155–62.

  33. 33.

    Zhang Y, Lu H, Dazin P, Kapila Y. Functional differences between integrin alpha4 and integrins alpha5/alphav in modulating the motility of human oral squamous carcinoma cells in response to the V region and heparin-binding domain of fibronectin. Exp Cell Res. 2004;295:48–58.

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This work was supported by Institut National du Cancer (INCA) grants (2013-070 and 2013-1-MELA-04) and the “Association pour la Recherche sur le Cancer”, Equipe labellisée ARC 2015. Labex SIGNALIFE, ANR-11 LABEX-0028-01, la Ville de Nice. NEH is a recipient of “Fondation pour la Recherche Medicale” and LABEX. NH is a recipient of the ARC. We thank Dr. Emmanuel Lemichez for providing the HACE1 expression vector, Dr. Cory Hu for providing the GST-HACE1 plasmid and Pr. Alessi for providing the K27R plasmid. Frédéric Rénier, Centre Méditerranéen de Médecine Moléculaire, Bioinformatic Unit and Nicolas Nottet are supported by The Cancéropôle PACA. The authors thank the C3M animal and imaging facilities.

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Author notes

  1. Edited by S. Fulda


  1. Biology and pathologies of melanocytes, Team 1, Inserm U1065, Equipe labellisée ARC 2015, C3M, Université Nice Côte d’Azur, Nice, France

    • Najla El-Hachem
    • , Nadia Habel
    • , Tanesha Naiken
    • , Hanene Bzioueche
    • , Yann Cheli
    • , Guillaume E. Beranger
    • , Emilie Jaune
    • , Florian Rouaud
    • , Frédéric Reinier
    • , Céline Gaudel
    • , Corine Bertolotto
    •  & Robert Ballotti
  2. CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, Université Nice Côte d’Azur, Sophia Antipolis, France

    • Nicolas Nottet
  3. Inserm U1065, Team 2, C3M, Université Nice Côte d’Azur, Nice, France

    • Pascale Colosetti


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Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Robert Ballotti.

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