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Differential apoptotic activities of wild-type FOXL2 and the adult-type granulosa cell tumor-associated mutant FOXL2 (C134W)

Abstract

Some mutations in FOXL2 result in premature ovarian failure accompanied by blepharophimosis, ptosis, epicanthus inversus syndrome type I disease, and FOXL2-null mice exhibit developmental defects in granulosa cells. Recently, FOXL2 c.402C>G, a new somatic mutation that leads to a p.C134W change, was found in the majority of adult-type ovarian granulosa cell tumors (GCTs). In this study, we investigated the possible mechanisms by which the C134W mutation contributes to the development of GCTs. Wild-type (WT) and mutant FOXL2 displayed differential apoptotic activities. Specifically, WT FOXL2 induced significant granulosa cell death, but the mutant exhibited minimal cell death. The FOXL2-induced apoptotic response was greatly dependent on caspase 8, BID and BAK because the depletion of any of these three proteins inhibited FOXL2 from eliciting the full apoptotic response. Activation of caspase 8 and subsequent increased production of truncated BID, and oligomerization of BAK, and release of cytochrome c were all associated with the apoptosis induced by WT FOXL2 expression. In contrast, the mutant FOXL2 was unable to elicit the full array of apoptotic signaling responses. In addition, we found differential TNF-R1 (tumor necrosis factor-receptor 1) and Fas (CD95/APO-1) upregulation between the WT and the mutant, and the silencing of TNF-R1 or Fas and the blockage of the death signaling mediated by TNF-R1 or Fas using TNF-Fc or Fas-Fc, respectively, resulted in significant attenuations of FOXL2-induced apoptosis. Moreover, granulosa cells that expressed either WT FOXL2 or mutant exhibited distinct cell death sensitivities on activation of death receptors and deprivation of serum. Thus, the differential activities of FOXL2 and its mutant may partially account for the pathophysiology of GCT development.

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References

  • Arnoult D . (2008). Apoptosis-associated mitochondrial outer membrane permeabilization assays. Methods 44: 229–234.

    Article  CAS  PubMed  Google Scholar 

  • Benayoun BA, Caburet S, Dipietromaria A, Georges A, D'Haene B, Pandaranayaka PJ et al. (2010). Functional exploration of the adult ovarian granulosa cell tumor-associated somatic FOXL2 mutation p.Cys134Trp (c.402C>G). PLoS One 5: e8789.

    Article  PubMed  PubMed Central  Google Scholar 

  • Chang DW, Xing Z, Pan Y, Algeciras-Schimnich A, Barnhart BC, Yaish-Ohad S et al. (2002). c-FLIP(L) is a dual function regulator for caspase-8 activation and CD95-mediated apoptosis. EMBO J 21: 3704–3714.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chipuk JE, Bouchier-Hayes L, Green DR . (2006). Mitochondrial outer membrane permeabilization during apoptosis: the innocent bystander scenario. Cell Death Differ 13: 1396–1402.

    Article  CAS  PubMed  Google Scholar 

  • Crisponi L, Deiana M, Loi A, Chiappe F, Uda M, Amati P et al. (2001). The putative forkhead transcription factor FOXL2 is mutated in blepharophimosis/ptosis/epicanthus inversus syndrome. Nat Genet 27: 159–166.

    Article  CAS  PubMed  Google Scholar 

  • Dewson G, Kratina T, Sim HW, Puthalakath H, Adams JM, Colman PM et al. (2008). To trigger apoptosis, Bak exposes its BH3 domain and homodimerizes via BH3:groove interactions. Mol Cell 30: 369–380.

    Article  CAS  PubMed  Google Scholar 

  • Ganju N, Eastman A . (2003). Zinc inhibits Bax and Bak activation and cytochrome c release induced by chemical inducers of apoptosis but not by death-receptor-initiated pathways. Cell Death Differ 10: 652–661.

    Article  CAS  PubMed  Google Scholar 

  • Green DR, Kroemer G . (2005). Pharmacological manipulation of cell death: clinical applications in sight? J Clin Invest 115: 2610–2617.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guicciardi ME, Gores GJ . (2009). Life and death by death receptors. FASEB J 23: 1625–1637.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hail Jr N . (2005). Mitochondria: a novel target for the chemoprevention of cancer. Apoptosis 10: 687–705.

    Article  CAS  PubMed  Google Scholar 

  • Jabbour AM, Heraud JE, Daunt CP, Kaufmann T, Sandow J, O'Reilly LA et al. (2009). Puma indirectly activates Bax to cause apoptosis in the absence of Bid or Bim. Cell Death Differ 16: 555–563.

    Article  CAS  PubMed  Google Scholar 

  • Kang BP, Frencher S, Reddy V, Kessler A, Malhotra A, Meggs LG . (2003). High glucose promotes mesangial cell apoptosis by oxidant-dependent mechanism. Am J Physiol Renal Physiol 284: F455–F466.

    Article  CAS  PubMed  Google Scholar 

  • Kim MS, Hur SY, Yoo NJ, Lee SH . (2010). Mutational analysis of FOXL2 codon 134 in granulosa cell tumour of ovary and other human cancers. J Pathol 221: 147–152.

    Article  CAS  PubMed  Google Scholar 

  • Kornblau SM, Singh N, Qiu Y, Chen W, Zhang N, Coombes KR . (2010). Highly phosphorylated FOXO3A is an adverse prognostic factor in acute myeloid leukemia. Clin Cancer Res 16: 1865–1874.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee K, Pisarska MD, Ko JJ, Kang Y, Yoon S, Ryou SM et al. (2005). Transcriptional factor FOXL2 interacts with DP103 and induces apoptosis. Biochem Biophys Res Commun 336: 876–881.

    Article  CAS  PubMed  Google Scholar 

  • Li J, Yuan J . (2008). Caspases in apoptosis and beyond. Oncogene 27: 6194–6206.

    Article  CAS  PubMed  Google Scholar 

  • Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES et al. (1997). Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489.

    Article  CAS  PubMed  Google Scholar 

  • Li P, Nijhawan D, Wang X . (2004). Mitochondrial activation of apoptosis. Cell 116: S57–S59.

    Article  CAS  PubMed  Google Scholar 

  • Mahmood Z, Shukla Y . (2010). Death receptors: targets for cancer therapy. Exp Cell Res 316: 887–899.

    Article  CAS  PubMed  Google Scholar 

  • Muzio M . (1998). Signalling by proteolysis: death receptors induce apoptosis. Int J Clin Lab Res 28: 141–147.

    Article  CAS  PubMed  Google Scholar 

  • Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J et al. (1996). FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85: 817–827.

    Article  CAS  PubMed  Google Scholar 

  • Naka K, Hoshii T, Muraguchi T, Tadokoro Y, Ooshio T, Kondo Y et al. (2010). TGF-beta-FOXO signalling maintains leukaemia-initiating cells in chronic myeloid leukaemia. Nature 463: 676–680.

    Article  CAS  PubMed  Google Scholar 

  • Nishi Y, Yanase T, Mu Y, Oba K, Ichino I, Saito M et al. (2001). Establishment and characterization of a steroidogenic human granulosa-like tumor cell line, KGN, that expresses functional follicle-stimulating hormone receptor. Endocrinology 142: 437–445.

    Article  CAS  PubMed  Google Scholar 

  • Obexer P, Geiger K, Ambros PF, Meister B, Ausserlechner MJ . (2007). FKHRL1-mediated expression of Noxa and Bim induces apoptosis via the mitochondria in neuroblastoma cells. Cell Death Differ 14: 534–547.

    Article  CAS  PubMed  Google Scholar 

  • Paik JH, Kollipara R, Chu G, Ji H, Xiao Y, Ding Z et al. (2007). FoxOs are lineage-restricted redundant tumor suppressors and regulate endothelial cell homeostasis. Cell 128: 309–323.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Park M, Shin E, Won M, Kim JH, Go H, Kim HL et al. (2010). FOXL2 interacts with steroidogenic factor-1 (SF-1) and represses SF-1-induced CYP17 transcription in granulosa cells. Mol Endocrinol 24: 1024–1036.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Perez D, White E . (2000). TNF-alpha signals apoptosis through a bid-dependent conformational change in Bax that is inhibited by E1B 19K. Mol Cell 6: 53–63.

    Article  CAS  PubMed  Google Scholar 

  • Pisarska MD, Bae J, Klein C, Hsueh AJ . (2004). Forkhead l2 is expressed in the ovary and represses the promoter activity of the steroidogenic acute regulatory gene. Endocrinology 145: 3424–3433.

    Article  CAS  PubMed  Google Scholar 

  • Schrader KA, Gorbatcheva B, Senz J, Heravi-Moussavi A, Melnyk N, Salamanca C et al. (2009). The specificity of the FOXL2 c.402C>G somatic mutation: a survey of solid tumors. PLoS One 4: e7988.

    Article  PubMed  PubMed Central  Google Scholar 

  • Schumer ST, Cannistra SA . (2003). Granulosa cell tumor of the ovary. J Clin Oncol 21: 1180–1189.

    Article  PubMed  Google Scholar 

  • Shah SP, Kobel M, Senz J, Morin RD, Clarke BA, Wiegand KC et al. (2009). Mutation of FOXL2 in granulosa-cell tumors of the ovary. N Engl J Med 360: 2719–2729.

    Article  CAS  PubMed  Google Scholar 

  • Uda M, Ottolenghi C, Crisponi L, Garcia JE, Deiana M, Kimber W et al. (2004). Foxl2 disruption causes mouse ovarian failure by pervasive blockage of follicle development. Hum Mol Genet 13: 1171–1181.

    Article  CAS  PubMed  Google Scholar 

  • Wei MC, Lindsten T, Mootha VK, Weiler S, Gross A, Ashiya M et al. (2000). tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrome c. Genes Dev 14: 2060–2071.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yin XM, Wang K, Gross A, Zhao Y, Zinkel S, Klocke B et al. (1999). Bid-deficient mice are resistant to Fas-induced hepatocellular apoptosis. Nature 400: 886–891.

    Article  CAS  PubMed  Google Scholar 

  • Zou H, Li Y, Liu X, Wang X . (1999). An APAF-1.cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J Biol Chem 274: 11549–11556.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Basic Science Research Program (2009-0066320) and the Priority Research Centers Program (2009-0093821) through the National Research Foundation of Korea (NRF) of the Ministry of Education, Science and Technology and by a grant (A084923) from the Korea Healthcare Technology R&D Project from the Ministry of Health, Welfare and Family Affairs.

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Correspondence to J Bae.

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Kim, JH., Yoon, S., Park, M. et al. Differential apoptotic activities of wild-type FOXL2 and the adult-type granulosa cell tumor-associated mutant FOXL2 (C134W). Oncogene 30, 1653–1663 (2011). https://doi.org/10.1038/onc.2010.541

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