Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

MLKL in cancer: more than a necroptosis regulator


Mixed lineage kinase domain-like protein (MLKL) emerged as executioner of necroptosis, a RIPK3-dependent form of regulated necrosis. Cell death evasion is one of the hallmarks of cancer. Besides apoptosis, some cancers suppress necroptosis-associated mechanisms by for example epigenetic silencing of RIPK3 expression. Conversely, necroptosis-elicited inflammation by cancer cells can fuel tumor growth. Recently, necroptosis-independent functions of MLKL were unraveled in receptor internalization, ligand-receptor degradation, endosomal trafficking, extracellular vesicle formation, autophagy, nuclear functions, axon repair, neutrophil extracellular trap (NET) formation, and inflammasome regulation. Little is known about the precise role of MLKL in cancer and whether some of these functions are involved in cancer development and metastasis. Here, we discuss current knowledge and controversies on MLKL, its structure, necroptosis-independent functions, expression, mutations, and its potential role as a pro- or anti-cancerous factor. Analysis of MLKL expression patterns reveals that MLKL is upregulated by type I/II interferon, conditions of inflammation, and tissue injury. Overall, MLKL may affect cancer development and metastasis through necroptosis-dependent and -independent functions.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: Multiple stimuli lead to necroptosis.
Fig. 2: Structural domains of MLKL and their functions.
Fig. 3: RIPK3 and MLKL genes are differentially expressed in tissues during homeostasis.
Fig. 4: Ripk3 and Mlkl are differentially expressed in tissues during homeostasis and during perturbations.
Fig. 5: Transcriptional regulation of MLKL/Mlkl.
Fig. 6: High variation in MLKL/Mlkl expression between different types of cancer.


  1. 1.

    Choi ME, Price DR, Ryter SW, Choi AMK. Necroptosis: a crucial pathogenic mediator of human disease. JCI Insight. 2019;4:e128834.

    PubMed Central  Google Scholar 

  2. 2.

    Sun L, Wang H, Wang Z, He S, Chen S, Liao D, et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell. 2012;148:213–27.

    CAS  PubMed  Google Scholar 

  3. 3.

    Zhao J, Jitkaew S, Cai Z, Choksi S, Li Q, Luo J, et al. Mixed lineage kinase domain-like is a key receptor interacting protein 3 downstream component of TNF-induced necrosis. Proc Natl Acad Sci USA. 2012;109:5322–7.

    CAS  PubMed  Google Scholar 

  4. 4.

    Pasparakis M, Vandenabeele P. Necroptosis and its role in inflammation. Nature. 2015;517:311–20.

    CAS  PubMed  Google Scholar 

  5. 5.

    Delanghe T, Dondelinger Y, Bertrand MJM. RIPK1 kinase-dependent death: a symphony of phosphorylation events. Trends Cell Biol. 2020;30:189–200.

    CAS  PubMed  Google Scholar 

  6. 6.

    Vercammen D, Beyaert R, Denecker G, Goossens V, Van Loo G, Declercq W, et al. Inhibition of caspases increases the sensitivity of L929 cells to necrosis mediated by tumor necrosis factor. J Exp Med. 1998;187:1477–85.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Newton K, Wickliffe KE, Dugger DL, Maltzman A, Roose-Girma M, Dohse M, et al. Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis. Nature. 2019;574:428–31.

    CAS  PubMed  Google Scholar 

  8. 8.

    Wallach D, Kang TB, Dillon CP, Green DR. Programmed necrosis in inflammation: toward identification of the effector molecules. Science. 2016;352:aaf2154.

    PubMed  Google Scholar 

  9. 9.

    Su Z, Yang Z, Xie L, DeWitt JP, Chen Y. Cancer therapy in the necroptosis era. Cell Death Differ. 2016;23:748–56.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. 10.

    Najafov A, Chen H, Yuan J. Necroptosis and cancer. Trends Cancer. 2017;3:294–301.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Gong Y, Fan Z, Luo G, Yang C, Huang Q, Fan K, et al. The role of necroptosis in cancer biology and therapy. Mol Cancer. 2019;18:100.

    PubMed  PubMed Central  Google Scholar 

  12. 12.

    Zhu F, Zhang W, Yang T, He SD. Complex roles of necroptosis in cancer. J Zhejiang Univ Sci B. 2019;20:399–413.

    PubMed  PubMed Central  Google Scholar 

  13. 13.

    Yatim N, Jusforgues-Saklani H, Orozco S, Schulz O, Barreira da Silva R, Reis e Sousa C, et al. RIPK1 and NF-kappaB signaling in dying cells determines cross-priming of CD8(+) T cells. Science. 2015;350:328–34.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Aaes TL, Kaczmarek A, Delvaeye T, De Craene B, De Koker S, Heyndrickx L, et al. Vaccination with necroptotic cancer cells induces efficient anti-tumor immunity. Cell Rep. 2016;15:274–87.

    CAS  PubMed  Google Scholar 

  15. 15.

    Yatim N, Cullen S, Albert ML. Dying cells actively regulate adaptive immune responses. Nat Rev Immunol. 2017;17:262–75.

    CAS  PubMed  Google Scholar 

  16. 16.

    Ying Z, Pan C, Shao T, Liu L, Li L, Guo D, et al. Mixed lineage kinase domain-like protein MLKL breaks down myelin following nerve injury. Mol Cell. 2018;72:457–68 e5.

    CAS  PubMed  Google Scholar 

  17. 17.

    Koo GB, Morgan MJ, Lee DG, Kim WJ, Yoon JH, Koo JS, et al. Methylation-dependent loss of RIP3 expression in cancer represses programmed necrosis in response to chemotherapeutics. Cell Res. 2015;25:707–25.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Lalaoui N, Brumatti G. Relevance of necroptosis in cancer. Immunol Cell Biol. 2017;95:137–45.

    CAS  PubMed  Google Scholar 

  19. 19.

    Petrie EJ, Hildebrand JM, Murphy JM. Insane in the membrane: a structural perspective of MLKL function in necroptosis. Immunol Cell Biol. 2017;95:152–9.

    CAS  PubMed  Google Scholar 

  20. 20.

    Hildebrand JM, Tanzer MC, Lucet IS, Young SN, Spall SK, Sharma P, et al. Activation of the pseudokinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death. Proc Natl Acad Sci USA. 2014;111:15072–7.

    CAS  PubMed  Google Scholar 

  21. 21.

    Chen X, Li W, Ren J, Huang D, He WT, Song Y, et al. Translocation of mixed lineage kinase domain-like protein to plasma membrane leads to necrotic cell death. Cell Res. 2014;24:105–21.

    CAS  PubMed  Google Scholar 

  22. 22.

    Tanzer MC, Matti I, Hildebrand JM, Young SN, Wardak A, Tripaydonis A, et al. Evolutionary divergence of the necroptosis effector MLKL. Cell Death Differ. 2016;23:1185–97.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Dondelinger Y, Declercq W, Montessuit S, Roelandt R, Goncalves A, Bruggeman I, et al. MLKL compromises plasma membrane integrity by binding to phosphatidylinositol phosphates. Cell Rep. 2014;7:971–81.

    CAS  PubMed  Google Scholar 

  24. 24.

    Wang H, Sun L, Su L, Rizo J, Liu L, Wang LF, et al. Mixed lineage kinase domain-like protein MLKL causes necrotic membrane disruption upon phosphorylation by RIP3. Mol Cell. 2014;54:133–46.

    CAS  PubMed  Google Scholar 

  25. 25.

    Boudeau J, Miranda-Saavedra D, Barton GJ, Alessi DR. Emerging roles of pseudokinases. Trends Cell Biol. 2006;16:443–52.

    CAS  PubMed  Google Scholar 

  26. 26.

    Xie T, Peng W, Yan C, Wu J, Gong X, Shi Y. Structural insights into RIP3-mediated necroptotic signaling. Cell Rep. 2013;5:70–8.

    CAS  PubMed  Google Scholar 

  27. 27.

    Grootjans S, Vanden Berghe T, Vandenabeele P. Initiation and execution mechanisms of necroptosis: an overview. Cell Death Differ. 2017;24:1184–95.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Rodriguez DA, Weinlich R, Brown S, Guy C, Fitzgerald P, Dillon CP, et al. Characterization of RIPK3-mediated phosphorylation of the activation loop of MLKL during necroptosis. Cell Death Differ. 2016;23:76–88.

    CAS  PubMed  Google Scholar 

  29. 29.

    Petrie EJ, Sandow JJ, Jacobsen AV, Smith BJ, Griffin MDW, Lucet IS, et al. Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis. Nat Commun. 2018;9:2422.

    PubMed  PubMed Central  Google Scholar 

  30. 30.

    Yoon S, Kovalenko A, Bogdanov K, Wallach D. MLKL, the protein that mediates necroptosis, also regulates endosomal trafficking and extracellular vesicle generation. Immunity 2017;47:51–65 e7.

    CAS  PubMed  Google Scholar 

  31. 31.

    Davies KA, Fitzgibbon C, Young SN, Garnish SE, Yeung W, Coursier D, et al. Distinct pseudokinase domain conformations underlie divergent activation mechanisms among vertebrate MLKL orthologues. Nat Commun. 2020;11:3060.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. 32.

    Murphy JM, Czabotar PE, Hildebrand JM, Lucet IS, Zhang JG, Alvarez-Diaz S, et al. The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. Immunity. 2013;39:443–53.

    CAS  PubMed  Google Scholar 

  33. 33.

    Petrie EJ, Czabotar PE, Murphy JM. The structural basis of necroptotic cell death signaling. Trends Biochem Sci. 2019;44:53–63.

    CAS  PubMed  Google Scholar 

  34. 34.

    Rubbelke M, Fiegen D, Bauer M, Binder F, Hamilton J, King J, et al. Locking mixed-lineage kinase domain-like protein in its auto-inhibited state prevents necroptosis. Proc Natl Acad Sci USA. 2020;117:33272–81.

    PubMed  Google Scholar 

  35. 35.

    Tanzer MC, Tripaydonis A, Webb AI, Young SN, Varghese LN, Hall C, et al. Necroptosis signalling is tuned by phosphorylation of MLKL residues outside the pseudokinase domain activation loop. Biochem J. 2015;471:255–65.

    CAS  PubMed  Google Scholar 

  36. 36.

    Najafov A, Mookhtiar AK, Luu HS, Ordureau A, Pan H, Amin PP, et al. TAM kinases promote necroptosis by regulating oligomerization of MLKL. Mol Cell. 2019;75:457–68 e4.

    CAS  PubMed  Google Scholar 

  37. 37.

    Hildebrand JM, Kauppi M, Majewski IJ, Liu Z, Cox AJ, Miyake S, et al. A missense mutation in the MLKL brace region promotes lethal neonatal inflammation and hematopoietic dysfunction. Nat Commun. 2020;11:3150.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Huang D, Zheng X, Wang ZA, Chen X, He WT, Zhang Y, et al. The MLKL channel in necroptosis is an octamer formed by tetramers in a dyadic process. Mol Cell Biol. 2017;37:e00497–16.

    PubMed  PubMed Central  Google Scholar 

  39. 39.

    Quarato G, Guy CS, Grace CR, Llambi F, Nourse A, Rodriguez DA, et al. Sequential engagement of distinct MLKL phosphatidylinositol-binding sites executes necroptosis. Mol Cell. 2016;61:589–601.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Dovey CM, Diep J, Clarke BP, Hale AT, McNamara DE, Guo H, et al. MLKL requires the inositol phosphate code to execute necroptosis. Mol Cell. 2018;70:936–48 e7.

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Jing L, Song F, Liu Z, Li J, Wu B, Fu Z, et al. MLKL-PITPalpha signaling-mediated necroptosis contributes to cisplatin-triggered cell death in lung cancer A549 cells. Cancer Lett. 2018;414:136–46.

    CAS  PubMed  Google Scholar 

  42. 42.

    Liu S, Liu H, Johnston A, Hanna-Addams S, Reynoso E, Xiang Y, et al. MLKL forms disulfide bond-dependent amyloid-like polymers to induce necroptosis. Proc Natl Acad Sci USA. 2017;114:E7450–E9.

    CAS  PubMed  Google Scholar 

  43. 43.

    Li J, McQuade T, Siemer AB, Napetschnig J, Moriwaki K, Hsiao YS, et al. The RIP1/RIP3 necrosome forms a functional amyloid signaling complex required for programmed necrosis. Cell. 2012;150:339–50.

    CAS  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Gong YN, Guy C, Olauson H, Becker JU, Yang M, Fitzgerald P, et al. ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences. Cell. 2017;169:286–300 e16.

    CAS  PubMed  PubMed Central  Google Scholar 

  45. 45.

    Guo H, Kaiser WJ. ESCRTing necroptosis. Cell. 2017;169:186–7.

    CAS  PubMed  Google Scholar 

  46. 46.

    van der Leun AM, Thommen DS, Schumacher TN. CD8(+) T cell states in human cancer: insights from single-cell analysis. Nat Rev Cancer. 2020;20:218–32.

    PubMed  PubMed Central  Google Scholar 

  47. 47.

    Do HTT, Lee CH, Cho J. Chemokines and their receptors: multifaceted roles in cancer progression and potential value as cancer prognostic markers. Cancers. 2020;12:287.

    CAS  PubMed Central  Google Scholar 

  48. 48.

    Dhawan P, Richmond A. Role of CXCL1 in tumorigenesis of melanoma. J Leukoc Biol. 2002;72:9–18.

    CAS  PubMed  PubMed Central  Google Scholar 

  49. 49.

    Acharyya S, Oskarsson T, Vanharanta S, Malladi S, Kim J, Morris PG, et al. A CXCL1 paracrine network links cancer chemoresistance and metastasis. Cell. 2012;150:165–78.

    CAS  PubMed  PubMed Central  Google Scholar 

  50. 50.

    Tokunaga R, Zhang W, Naseem M, Puccini A, Berger MD, Soni S, et al. CXCL9, CXCL10, CXCL11/CXCR3 axis for immune activation—a target for novel cancer therapy. Cancer Treat Rev. 2018;63:40–7.

    CAS  Google Scholar 

  51. 51.

    Doron H, Amer M, Ershaid N, Blazquez R, Shani O, Lahav TG, et al. InflammatorY Activation of Astrocytes Facilitates Melanoma Brain Tropism via the CXCL10-CXCR3 signaling axis. Cell Rep. 2019;28:1785–98 e6.

    CAS  PubMed  Google Scholar 

  52. 52.

    Zhu G, Yan HH, Pang Y, Jian J, Achyut BR, Liang X, et al. CXCR3 as a molecular target in breast cancer metastasis: inhibition of tumor cell migration and promotion of host anti-tumor immunity. Oncotarget. 2015;6:43408–19.

    PubMed  PubMed Central  Google Scholar 

  53. 53.

    Frank D, Vaux DL, Murphy JM, Vince JE, Lindqvist LM. Activated MLKL attenuates autophagy following its translocation to intracellular membranes. J Cell Sci. 2019;132:jcs220996.

    CAS  PubMed  Google Scholar 

  54. 54.

    Wu X, Poulsen KL, Sanz-Garcia C, Huang E, McMullen MR, Roychowdhury S, et al. MLKL-dependent signaling regulates autophagic flux in a murine model of non-alcohol-associated fatty liver and steatohepatitis. J Hepatol. 2020;73:616–27.

    CAS  PubMed  Google Scholar 

  55. 55.

    Santana-Codina N, Mancias JD, Kimmelman AC. The role of autophagy in cancer. Annu Rev Cancer Biol. 2017;1:19–39.

    PubMed  PubMed Central  Google Scholar 

  56. 56.

    Yun CW, Lee SH. The roles of autophagy in cancer. Int J Mol Sci. 2018;19:3466.

    PubMed Central  Google Scholar 

  57. 57.

    Mulcahy Levy JM, Thorburn A. Autophagy in cancer: moving from understanding mechanism to improving therapy responses in patients. Cell Death Differ. 2020;27:843–57.

    PubMed  Google Scholar 

  58. 58.

    Liu S, Li Y, Choi HMC, Sarkar C, Koh EY, Wu J, et al. Lysosomal damage after spinal cord injury causes accumulation of RIPK1 and RIPK3 proteins and potentiation of necroptosis. Cell Death Dis. 2018;9:476.

    PubMed  PubMed Central  Google Scholar 

  59. 59.

    Moriwaki K, Balaji S, Bertin J, Gough PJ, Chan FK. Distinct kinase-independent role of RIPK3 in CD11c(+) mononuclear phagocytes in cytokine-induced tissue repair. Cell Rep. 2017;18:2441–51.

    CAS  PubMed  PubMed Central  Google Scholar 

  60. 60.

    Zhou S, Zhang W, Cai G, Ding Y, Wei C, Li S, et al. Myofiber necroptosis promotes muscle stem cell proliferation via releasing Tenascin-C during regeneration. Cell Res. 2020;30:1063–77.

    PubMed  PubMed Central  Google Scholar 

  61. 61.

    Yoon S, Bogdanov K, Kovalenko A, Wallach D. Necroptosis is preceded by nuclear translocation of the signaling proteins that induce it. Cell Death Differ. 2016;23:253–60.

    CAS  PubMed  Google Scholar 

  62. 62.

    Weber K, Roelandt R, Bruggeman I, Estornes Y, Vandenabeele P. Nuclear RIPK3 and MLKL contribute to cytosolic necrosome formation and necroptosis. Commun Biol. 2018;1:6.

    PubMed  PubMed Central  Google Scholar 

  63. 63.

    Cai F, Wang JL, Wu YL, Hu YW, Wang Q. Mixed lineage kinase domain-like protein promotes human monocyte cell adhesion to human umbilical vein endothelial cells via upregulation of intercellular adhesion molecule-1 expression. Med Sci Monit. 2020;26:e924242.

    CAS  PubMed  PubMed Central  Google Scholar 

  64. 64.

    Dai J, Zhang C, Guo L, He H, Jiang K, Huang Y, et al. A necroptotic-independent function of MLKL in regulating endothelial cell adhesion molecule expression. Cell Death Dis. 2020;11:282.

    CAS  PubMed  PubMed Central  Google Scholar 

  65. 65.

    Janiszewska M, Primi MC, Izard T. Cell adhesion in cancer: beyond the migration of single cells. J Biol Chem. 2020;295:2495–505.

    CAS  PubMed  PubMed Central  Google Scholar 

  66. 66.

    Conos SA, Chen KW, De Nardo D, Hara H, Whitehead L, Nunez G, et al. Active MLKL triggers the NLRP3 inflammasome in a cell-intrinsic manner. Proc Natl Acad Sci USA. 2017;114:E961–E9.

    CAS  PubMed  Google Scholar 

  67. 67.

    Gutierrez KD, Davis MA, Daniels BP, Olsen TM, Ralli-Jain P, Tait SW, et al. MLKL activation triggers NLRP3-mediated processing and release of IL-1beta Independently of gasdermin-D. J Immunol. 2017;198:2156–64.

    CAS  PubMed  PubMed Central  Google Scholar 

  68. 68.

    D’Cruz AA, Speir M, Bliss-Moreau M, Dietrich S, Wang S, Chen AA, et al. The pseudokinase MLKL activates PAD4-dependent NET formation in necroptotic neutrophils. Sci Signal. 2018;11:eaao1716.

    PubMed  PubMed Central  Google Scholar 

  69. 69.

    Teijeira A, Garasa S, Gato M, Alfaro C, Migueliz I, Cirella A, et al. CXCR1 and CXCR2 chemokine receptor agonists produced by tumors induce neutrophil extracellular traps that interfere with immune cytotoxicity. Immunity. 2020;52:856–71 e8.

    CAS  PubMed  Google Scholar 

  70. 70.

    Chessler AD, Unnikrishnan M, Bei AK, Daily JP, Burleigh BA. Trypanosoma cruzi triggers an early type I IFN response in vivo at the site of intradermal infection. J Immunol. 2009;182:2288–96.

    CAS  PubMed  Google Scholar 

  71. 71.

    Alvarez-Diaz S, Preaudet A, Samson AL, Nguyen PM, Fung KY, Garnham AL, et al. Necroptosis is dispensable for the development of inflammation-associated or sporadic colon cancer in mice. Cell Death Differ. 2020.

  72. 72.

    McComb S, Cessford E, Alturki NA, Joseph J, Shutinoski B, Startek JB, et al. Type-I interferon signaling through ISGF3 complex is required for sustained Rip3 activation and necroptosis in macrophages. Proc Natl Acad Sci USA. 2014;111:E3206–13.

    CAS  PubMed  Google Scholar 

  73. 73.

    Robinson N, McComb S, Mulligan R, Dudani R, Krishnan L, Sad S. Type I interferon induces necroptosis in macrophages during infection with Salmonella enterica serovar Typhimurium. Nat Immunol. 2012;13:954–62.

    CAS  PubMed  PubMed Central  Google Scholar 

  74. 74.

    Legarda D, Justus SJ, Ang RL, Rikhi N, Li W, Moran TM, et al. CYLD proteolysis protects macrophages from TNF-mediated auto-necroptosis induced by LPS and licensed by type I IFN. Cell Rep. 2016;15:2449–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  75. 75.

    Thapa RJ, Nogusa S, Chen P, Maki JL, Lerro A, Andrake M, et al. Interferon-induced RIP1/RIP3-mediated necrosis requires PKR and is licensed by FADD and caspases. Proc Natl Acad Sci USA. 2013;110:E3109–18.

    CAS  PubMed  Google Scholar 

  76. 76.

    Li Y, Guo X, Hu C, Du Y, Guo C, Di W, et al. Type I IFN operates pyroptosis and necroptosis during multidrug-resistant A. baumannii infection. Cell Death Differ. 2018;25:1304–18.

    CAS  PubMed  PubMed Central  Google Scholar 

  77. 77.

    Sarhan J, Liu BC, Muendlein HI, Weindel CG, Smirnova I, Tang AY, et al. Constitutive interferon signaling maintains critical threshold of MLKL expression to license necroptosis. Cell Death Differ. 2019;26:332–47.

    CAS  PubMed  Google Scholar 

  78. 78.

    Cerps SC, Menzel M, Mahmutovic Persson I, Bjermer L, Akbarshahi H, Uller L. Interferon-beta deficiency at asthma exacerbation promotes MLKL mediated necroptosis. Sci Rep. 2018;8:4248.

    PubMed  PubMed Central  Google Scholar 

  79. 79.

    Dillon CP, Weinlich R, Rodriguez DA, Cripps JG, Quarato G, Gurung P, et al. RIPK1 blocks early postnatal lethality mediated by caspase-8 and RIPK3. Cell. 2014;157:1189–202.

    CAS  PubMed  PubMed Central  Google Scholar 

  80. 80.

    Gunther C, He GW, Kremer AE, Murphy JM, Petrie EJ, Amann K, et al. The pseudokinase MLKL mediates programmed hepatocellular necrosis independently of RIPK3 during hepatitis. J Clin Investig. 2016;126:4346–60.

    PubMed  Google Scholar 

  81. 81.

    Lee SH, Kwon JY, Kim SY, Jung K, Cho ML. Interferon-gamma regulates inflammatory cell death by targeting necroptosis in experimental autoimmune arthritis. Sci Rep. 2017;7:10133.

    PubMed  PubMed Central  Google Scholar 

  82. 82.

    Cekay MJ, Roesler S, Frank T, Knuth AK, Eckhardt I, Fulda S. Smac mimetics and type II interferon synergistically induce necroptosis in various cancer cell lines. Cancer Lett. 2017;410:228–37.

    CAS  PubMed  Google Scholar 

  83. 83.

    Knuth AK, Rosler S, Schenk B, Kowald L, van Wijk SJL, Fulda S. Interferons transcriptionally up-regulate MLKL expression in cancer cells. Neoplasia. 2019;21:74–81.

    CAS  PubMed  Google Scholar 

  84. 84.

    Kursunel MA, Esendagli G. The untold story of IFN-gamma in cancer biology. Cytokine Growth Factor Rev. 2016;31:73–81.

    PubMed  Google Scholar 

  85. 85.

    Thibaut R, Bost P, Milo I, Cazaux M, Lemaitre F, Garcia Z, et al. Bystander IFN-gamma activity promotes widespread and sustained cytokine signaling altering the tumor microenvironment. Nat Cancer. 2020;1:302–14.

    PubMed  PubMed Central  Google Scholar 

  86. 86.

    Xiong Y, Li L, Zhang L, Cui Y, Wu C, Li H, et al. The bromodomain protein BRD4 positively regulates necroptosis via modulating MLKL expression. Cell Death Differ. 2019;26:1929–41.

    CAS  PubMed  PubMed Central  Google Scholar 

  87. 87.

    Guida N, Laudati G, Serani A, Mascolo L, Molinaro P, Montuori P, et al. The neurotoxicant PCB-95 by increasing the neuronal transcriptional repressor REST down-regulates caspase-8 and increases Ripk1, Ripk3 and MLKL expression determining necroptotic neuronal death. Biochem Pharm. 2017;142:229–41.

    CAS  PubMed  Google Scholar 

  88. 88.

    Sun SC. CYLD: a tumor suppressor deubiquitinase regulating NF-kappaB activation and diverse biological processes. Cell Death Differ. 2010;17:25–34.

    CAS  PubMed  PubMed Central  Google Scholar 

  89. 89.

    Forbes SA, Beare D, Bindal N, Bamford S, Ward S, Cole CG, et al. COSMIC: high-resolution cancer genetics using the catalogue of somatic mutations in cancer. Curr Protoc Hum Genet. 2016;91:10 1 1–1 37.

    Google Scholar 

  90. 90.

    Tate JG, Bamford S, Jubb HC, Sondka Z, Beare DM, Bindal N, et al. COSMIC: the catalogue of somatic mutations in cancer. Nucleic Acids Res. 2019;47:D941–D7.

    CAS  PubMed  Google Scholar 

  91. 91.

    Murphy JM, Lucet IS, Hildebrand JM, Tanzer MC, Young SN, Sharma P, et al. Insights into the evolution of divergent nucleotide-binding mechanisms among pseudokinases revealed by crystal structures of human and mouse MLKL. Biochem J. 2014;457:369–77.

    CAS  PubMed  Google Scholar 

  92. 92.

    Colbert LE, Fisher SB, Hardy CW, Hall WA, Saka B, Shelton JW, et al. Pronecrotic mixed lineage kinase domain-like protein expression is a prognostic biomarker in patients with early-stage resected pancreatic adenocarcinoma. Cancer 2013;119:3148–55.

    CAS  PubMed  PubMed Central  Google Scholar 

  93. 93.

    Seldon CS, Colbert LE, Hall WA, Fisher SB, Yu DS, Landry JC. Chromodomain-helicase-DNA binding protein 5, 7 and pronecrotic mixed lineage kinase domain-like protein serve as potential prognostic biomarkers in patients with resected pancreatic adenocarcinomas. World J Gastrointest Oncol. 2016;8:358–65.

    PubMed  PubMed Central  Google Scholar 

  94. 94.

    Li X, Guo J, Ding AP, Qi WW, Zhang PH, Lv J, et al. Association of mixed lineage kinase domain-like protein expression with prognosis in patients with colon cancer. Technol Cancer Res Treat. 2017;16:428–34.

    CAS  PubMed  Google Scholar 

  95. 95.

    Ertao Z, Jianhui C, Kang W, Zhijun Y, Hui W, Chuangqi C, et al. Prognostic value of mixed lineage kinase domain-like protein expression in the survival of patients with gastric caner. Tumour Biol. 2016;37:13679–85.

    PubMed  Google Scholar 

  96. 96.

    Li L, Yu S, Zang C. Low necroptosis process predicts poor treatment outcome of human papillomavirus positive cervical cancers by decreasing tumor-associated macrophages M1 polarization. Gynecol Obstet Investig. 2018;83:259–67.

    CAS  Google Scholar 

  97. 97.

    Ruan J, Mei L, Zhu Q, Shi G, Wang H. Mixed lineage kinase domain-like protein is a prognostic biomarker for cervical squamous cell cancer. Int J Clin Exp Pathol. 2015;8:15035–8.

    PubMed  PubMed Central  Google Scholar 

  98. 98.

    He L, Peng K, Liu Y, Xiong J, Zhu FF. Low expression of mixed lineage kinase domain-like protein is associated with poor prognosis in ovarian cancer patients. Onco Targets Ther. 2013;6:1539–43.

    CAS  PubMed  PubMed Central  Google Scholar 

  99. 99.

    Hockendorf U, Yabal M, Jost PJ. Killing AML: RIPK3 leads the way. Cell Cycle. 2017;16:3–4.

    PubMed  Google Scholar 

  100. 100.

    Hockendorf U, Yabal M, Herold T, Munkhbaatar E, Rott S, Jilg S, et al. RIPK3 restricts myeloid leukemogenesis by promoting cell death and differentiation of leukemia initiating cells. Cancer Cell. 2016;30:75–91.

    PubMed  Google Scholar 

  101. 101.

    Stoll G, Ma Y, Yang H, Kepp O, Zitvogel L, Kroemer G. Pro-necrotic molecules impact local immunosurveillance in human breast cancer. Oncoimmunology. 2017;6:e1299302.

    PubMed  PubMed Central  Google Scholar 

  102. 102.

    Nugues AL, El Bouazzati H, Hetuin D, Berthon C, Loyens A, Bertrand E, et al. RIP3 is downregulated in human myeloid leukemia cells and modulates apoptosis and caspase-mediated p65/RelA cleavage. Cell Death Dis. 2014;5:e1384.

    CAS  PubMed  PubMed Central  Google Scholar 

  103. 103.

    Hu B, Shi D, Lv X, Chen S, Huang Q, Xie M, et al. Prognostic and clinicopathological significance of MLKL expression in cancer patients: a meta-analysis. BMC Cancer. 2018;18:736.

    PubMed  PubMed Central  Google Scholar 

  104. 104.

    Aaes TL, Verschuere H, Kaczmarek A, Heyndrickx L, Wiernicki B, Delrue I, et al. Immunodominant AH1 antigen-deficient necroptotic, but not apoptotic, murine cancer cells induce antitumor protection. J Immunol. 2020;204:775–87.

    CAS  PubMed  Google Scholar 

  105. 105.

    Van Hoecke L, Van Lint S, Roose K, Van Parys A, Vandenabeele P, Grooten J, et al. Treatment with mRNA coding for the necroptosis mediator MLKL induces antitumor immunity directed against neo-epitopes. Nat Commun. 2018;9:3417.

    PubMed  PubMed Central  Google Scholar 

  106. 106.

    Van Hoecke L, Riederer S, Saelens X, Sutter G, Rojas JJ. Recombinant viruses delivering the necroptosis mediator MLKL induce a potent antitumor immunity in mice. Oncoimmunology. 2020;9:1802968.

    PubMed  PubMed Central  Google Scholar 

  107. 107.

    Sun D, Zhao L, Lin J, Zhao Y, Zheng Y. Cationic liposome co-encapsulation of SMAC mimetic and zVAD using a novel lipid bilayer fusion loaded with MLKL-pDNA for tumour inhibition in vivo. J Drug Target. 2018;26:45–54.

    CAS  PubMed  Google Scholar 

  108. 108.

    Yang H, Ma Y, Chen G, Zhou H, Yamazaki T, Klein C, et al. Contribution of RIP3 and MLKL to immunogenic cell death signaling in cancer chemotherapy. Oncoimmunology. 2016;5:e1149673.

    PubMed  PubMed Central  Google Scholar 

  109. 109.

    Seifert L, Werba G, Tiwari S, Giao Ly NN, Alothman S, Alqunaibit D, et al. The necrosome promotes pancreatic oncogenesis via CXCL1 and Mincle-induced immune suppression. Nature. 2016;532:245–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  110. 110.

    Seifert L, Miller G. Molecular pathways: the necrosome-a target for cancer therapy. Clin Cancer Res. 2017;23:1132–6.

    CAS  PubMed  Google Scholar 

  111. 111.

    Ando Y, Ohuchida K, Otsubo Y, Kibe S, Takesue S, Abe T, et al. Necroptosis in pancreatic cancer promotes cancer cell migration and invasion by release of CXCL5. PLoS One. 2020;15:e0228015.

    CAS  PubMed  PubMed Central  Google Scholar 

  112. 112.

    Liu XJ, Zhou M, Mei LY, Ruan JY, Hu Q, Peng J, et al. Key roles of necroptotic factors in promoting tumor growth. Oncotarget. 2016;7:22219–33.

    PubMed  PubMed Central  Google Scholar 

  113. 113.

    Dong Y, Sun Y, Huang Y, Dwarakanath B, Kong L, Lu JJ. Upregulated necroptosis-pathway-associated genes are unfavorable prognostic markers in low-grade glioma and glioblastoma multiforme. Transl Cancer Res. 2019;8:821–7.

    CAS  Google Scholar 

  114. 114.

    Li J, Huang S, Zeng L, Li K, Yang L, Gao S, et al. Necroptosis in head and neck squamous cell carcinoma: characterization of clinicopathological relevance and in vitro cell model. Cell Death Dis. 2020;11:391.

    CAS  PubMed  PubMed Central  Google Scholar 

  115. 115.

    Xin J, You D, Breslin P, Li J, Zhang J, Wei W, et al. Sensitizing acute myeloid leukemia cells to induced differentiation by inhibiting the RIP1/RIP3 pathway. Leukemia. 2017;31:1154–65.

    CAS  PubMed  Google Scholar 

  116. 116.

    Strilic B, Yang L, Albarran-Juarez J, Wachsmuth L, Han K, Muller UC, et al. Tumour-cell-induced endothelial cell necroptosis via death receptor 6 promotes metastasis. Nature. 2016;536:215–8.

    CAS  PubMed  Google Scholar 

  117. 117.

    Hanggi K, Vasilikos L, Valls AF, Yerbes R, Knop J, Spilgies LM, et al. RIPK1/RIPK3 promotes vascular permeability to allow tumor cell extravasation independent of its necroptotic function. Cell Death Dis. 2017;8:e2588.

    CAS  PubMed  PubMed Central  Google Scholar 

  118. 118.

    Jiao D, Cai Z, Choksi S, Ma D, Choe M, Kwon HJ, et al. Necroptosis of tumor cells leads to tumor necrosis and promotes tumor metastasis. Cell Res. 2018;28:868–70.

    CAS  PubMed  PubMed Central  Google Scholar 

  119. 119.

    Patel S, Webster JD, Varfolomeev E, Kwon YC, Cheng JH, Zhang J, et al. RIP1 inhibition blocks inflammatory diseases but not tumor growth or metastases. Cell Death Differ. 2020;27:161–75.

    CAS  PubMed  Google Scholar 

  120. 120.

    Dong Y, Sun Y, Huang Y, Fang X, Sun P, Dwarakanath B, et al. Depletion of MLKL inhibits invasion of radioresistant nasopharyngeal carcinoma cells by suppressing epithelial-mesenchymal transition. Ann Transl Med. 2019;7:741.

    CAS  PubMed  PubMed Central  Google Scholar 

  121. 121.

    Cai Z, Zhang A, Choksi S, Li W, Li T, Zhang XM, et al. Activation of cell-surface proteases promotes necroptosis, inflammation and cell migration. Cell Res. 2016;26:886–900.

    CAS  PubMed  PubMed Central  Google Scholar 

  122. 122.

    Han L, Lam EW, Sun Y. Extracellular vesicles in the tumor microenvironment: old stories, but new tales. Mol Cancer. 2019;18:59.

    PubMed  PubMed Central  Google Scholar 

  123. 123.

    Maacha S, Bhat AA, Jimenez L, Raza A, Haris M, Uddin S, et al. Extracellular vesicles-mediated intercellular communication: roles in the tumor microenvironment and anti-cancer drug resistance. Mol Cancer. 2019;18:55.

    PubMed  PubMed Central  Google Scholar 

  124. 124.

    Sheehan C, D’Souza-Schorey C. Tumor-derived extracellular vesicles: molecular parcels that enable regulation of the immune response in cancer. J Cell Sci. 2019;132:jcs235085.

    CAS  PubMed  PubMed Central  Google Scholar 

  125. 125.

    Snoderly HT, Boone BA, Bennewitz MF. Neutrophil extracellular traps in breast cancer and beyond: current perspectives on NET stimuli, thrombosis and metastasis, and clinical utility for diagnosis and treatment. Breast Cancer Res. 2019;21:145.

    CAS  PubMed  PubMed Central  Google Scholar 

  126. 126.

    Albrengues J, Shields MA, Ng D, Park CG, Ambrico A, Poindexter ME, et al. Neutrophil extracellular traps produced during inflammation awaken dormant cancer cells in mice. Science. 2018;361:eaao4227.

    PubMed  PubMed Central  Google Scholar 

  127. 127.

    Flemming A. Tumours use NETs as physical shields. Nat Rev Drug Discov. 2020;19:388.

    CAS  PubMed  Google Scholar 

  128. 128.

    Johnston A, Wang Z. Necroptosis: MLKL polymerization. J Nat Sci. 2018;4:e513.

    PubMed  PubMed Central  Google Scholar 

Download references


SM and JB obtained a PhD fellowship from the IWT (now FWO-SB). SM, JB, NT were paid by Methusalem. Research in the Vandenabeele group is supported by Flemish grants (EOS MODEL-IDI, FWO Grant 30826052), FWO research grants (G.0E04.16N, G.0C76.18N, G.0B71.18N, G.0B96.20N), Methusalem (BOF16/MET_V/007), iBOF20/IBF/039 ATLANTIS, Foundation against Cancer (FAF-F/2016/865, F/2020/1505), CRIG and GIGG consortia, and VIB.


Funding for this work is mentioned in the acknowledgements.

Author information




SM: concept, design of figures, performing analysis, writing. JB: correction of text and figures. RR: performing analysis, figures. PV: concept, design of figures, writing and correction of text. NT: concept, design of figures, writing and correction of text

Corresponding author

Correspondence to Peter Vandenabeele.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethics statement

This review did not require ethical approval.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Edited by F. Pentimalli

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Martens, S., Bridelance, J., Roelandt, R. et al. MLKL in cancer: more than a necroptosis regulator. Cell Death Differ 28, 1757–1772 (2021).

Download citation


Quick links