Abstract
Intestinal epithelial cell (IEC) death signalling through the Fas receptor is impaired in active ulcerative colitis (UC). This is possibly due to the activation of cytoprotective pathways resulting in limitation of the tissue injury secondary to inflammation. We hypothesized that inflammatory signalling like the nuclear factor (NF)-κB or mitogen activated protein kinase (MAPK) pathways could be involved in (a) the modification of Fas mediated apoptosis responses and (b) the regulation of the Fas receptor inhibitor cellular FLICE-like inhibitory protein (c-FLIP). Phospho-ERK was upregulated in IECs in active UC as well as IECs exposed to pro-inflammatory cytokines in vitro. Similarly, the short form of c-FLIP (c-FLIPS) was found to be upregulated in IECs from patients with active UC. c-FLIPS was the main splice variant found in both HT-29 cells and primary human IECs. Both splice variants were induced by TNF-α, IL-1β and IFN-γ, while IL-10 induced c-FLIPL expression; TNF-α also induced c-FLIPS in primary IECs. Inhibition of NF-κB, JNK and p38 pathways did not affect c-FLIP expression, whereas ERK inhibition by MEK1 RNA silencing and pharmacologic inhibitors decreased c-FLIPS expression. Similarly, ERK – but not NF-κB – inhibited Fas ligand and TNF-α-mediated apoptosis responses in both cell line experiments and primary IECs. The present study identifies the MEK-ERK pathway as a major regulator of apoptosis in IECs during flares of UC and an inducer of c-FLIPS. The results explain the resistance to receptor mediated epithelial apoptosis in active UC. Oncogenic c-FLIP could promote propagation of DNA-damaged IECs and contribute to cancer development in UC.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Begue B, Wajant H, Bambou JC, Dubuquoy L, Siegmund D, Beaulieu JF, Canioni D, Berrebi D, Brousse N, Desreumaux P et al. (2006) Implication of TNF-related apoptosis-inducing ligand in inflammatory intestinal epithelial lesions. Gastroenterology 130:1962–1974
Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9:799–809
Monteleone G, Caruso R, Fina D, Peluso I, Gioia V, Stolfi C, Fantini MC, Caprioli F, Tersigni R, Alessandroni L et al. (2006) Control of matrix metalloproteinase production in human intestinal fibroblasts by interleukin 21. Gut 55:1774–1780
Brand S (2009) Crohn’s disease: Th1, Th17 or both? The change of a paradigm: new immunological and genetic insights implicate Th17 cells in the pathogenesis of Crohn’s disease. Gut 58:1152–1167
Iimura M, Nakamura T, Shinozaki S, Iizuka B, Inoue Y, Suzuki S, Hayashi N (2000) Bax is downregulated in inflamed colonic mucosa of ulcerative colitis. Gut 47:228–235
Strauch ED, Bass BL, Rao JN, Vann JA, Wang JY (2003) NF-kappaB regulates intestinal epithelial cell and bile salt-induced migration after injury. Ann Surg 237:494–501
Egan LJ, Eckmann L, Greten FR, Chae S, Li ZW, Myhre GM, Robine S, Karin M, Kagnoff MF (2004) IkappaB-kinasebeta-dependent NF-kappaB activation provides radioprotection to the intestinal epithelium. Proc Natl Acad Sci USA 101:2452–2457
Karrasch T, Steinbrecher KA, Allard B, Baldwin AS, Jobin C (2006) Wound-induced p38MAPK-dependent histone H3 phosphorylation correlates with increased COX-2 expression in enterocytes. J Cell Physiol 207:809–815
Karrasch T, Jobin C (2009) Wound healing responses at the gastrointestinal epithelium: a close look at novel regulatory factors and investigative approaches. Z Gastroenterol 47:1221–1229
Seidelin JB, Nielsen OH (2008) Attenuated apoptosis response to Fas-ligand in active ulcerative colitis. Inflamm Bowel Dis 14:1623–1629
Hehlgans T, Pfeffer K (2005) The intriguing biology of the tumour necrosis factor/tumour necrosis factor receptor superfamily: players, rules and the games. Immunology 115:1–20
Tourneur L, Chiocchia G (2010) FADD: a regulator of life and death. Trends Immunol 31:260–269
Yu JW, Shi Y (2008) FLIP and the death effector domain family. Oncogene 27:6216–6227
Schroeder KW, Tremaine WJ, Ilstrup DM (1987) Coated oral 5-aminosalicylic acid therapy for mildly to moderately active ulcerative colitis. A randomized study. N Engl J Med 317:1625–1629
Langholz E (1999) Ulcerative colitis. An epidemiological study based on a regional inception cohort, with special reference to disease course and prognosis. Dan Med Bull 46:400–415
Seidelin JB, Horn T, Nielsen OH (2003) Simple and efficient method for isolation and cultivation of endoscopically obtained human colonocytes. Am J Physiol Gastrointest Liver Physiol 285:G1122–G1128
Seidelin JB, Vainer B, Andresen L, Nielsen OH (2007) Upregulation of cIAP2 in regenerating colonocytes in ulcerative colitis. Virchows Arch 415:1031–1038
Bharhani MS, Borojevic R, Basak S, Ho E, Zhou P, Croitoru K (2006) IL-10 protects mouse intestinal epithelial cells from Fas-induced apoptosis via modulating Fas expression and altering caspase-8 and FLIP expression. Am J Physiol Gastrointest Liver Physiol 291:G820-G829
Micheau O, Lens S, Gaide O, Alevizopoulos K, Tschopp J (2001) NF-kappaB signals induce the expression of c-FLIP. Mol Cell Biol 21:5299–5305
Ohori M, Kinoshita T, Okubo M, Sato K, Yamazaki A, Arakawa H, Nishimura S, Inamura N, Nakajima H, Neya M et al. (2005) Identification of a selective ERK inhibitor and structural determination of the inhibitor-ERK2 complex. Biochem Biophys Res Commun 336:357–363
Hommes D, van den BB, Plasse T, Bartelsman J, Xu C, Macpherson B, Tytgat G, Peppelenbosch M, van DS (2002) Inhibition of stress-activated MAP kinases induces clinical improvement in moderate to severe Crohn’s disease. Gastroenterology 122:7–14
Waetzig GH, Seegert D, Rosenstiel P, Nikolaus S, Schreiber S (2002) p38 mitogen-activated protein kinase is activated and linked to TNF-alpha signaling in inflammatory bowel disease. J Immunol 168:5342–5351
Zimmerman NP, Vongsa RA, Faherty SL, Salzman NH, Dwinell MB (2011) Targeted intestinal epithelial deletion of the chemokine receptor CXCR4 reveals important roles for extracellular-regulated kinase-1/2 in restitution. Laboratory Investigation 91:1040–1055
Yan F, John SK, Polk DB (2001) Kinase suppressor of Ras determines survival of intestinal epithelial cells exposed to tumor necrosis factor. Cancer Research 61:8668–8675
Yan F, Polk DB (2001) Kinase suppressor of ras is necessary for tumor necrosis factor alpha activation of extracellular signal-regulated kinase/mitogen-activated protein kinase in intestinal epithelial cells. Cancer Research 61:963–969
Wang WL, Prince CZ, Mou YS, Pollman MJ (2002) Notch3 signaling in vascular smooth muscle cells induces c-FLIP expression via ERK/MAPK activation—resistance to Fas ligand-induced apoptosis. Journal of Biological Chemistry 277:21723–21729
Seidelin JB, Jaattela M, Nielsen OH (2004) Continuous interferon-gamma or tumor necrosis factor-alpha exposure of enterocytes attenuates cell death responses. Cytokine 27:113–119
Todaro M, Zerilli M, Ricci-Vitiani L, Bini M, Perez AM, Maria FA, Miceli L, Condorelli G, Bonventre S, Di GG et al. (2006) Autocrine production of interleukin-4 and interleukin-10 is required for survival and growth of thyroid cancer cells. Cancer Res 66:1491–1499
Eslick J, Scatizzi JC, Albee L, Bickel E, Bradley K, Perlman H (2004) IL-4 and IL-10 inhibition of spontaneous monocyte apoptosis is associated with Flip upregulation. Inflammation 28:139–145
Santiago B, Galindo M, Palao G, Pablos JL (2004) Intracellular regulation of Fas-induced apoptosis in human fibroblasts by extracellular factors and cycloheximide. J Immunol 172:560–566
Llobet D, Eritja N, Domingo M, Bergada L, Mirantes C, Santacana M, Pallares J, Macia A, Yeramian A, Encinas M et al. (2011) KSR1 is overexpressed in endometrial carcinoma and regulates proliferation and TRAIL-induced apoptosis by modulating FLIP levels. Am J Pathol 178:1529–1543
Golks A, Brenner D, Krammer PH, Lavrik IN (2006) The c-FLIP-NH2 terminus (p22-FLIP) induces NF-kappaB activation. J Exp Med 203:1295–1305
Caprioli F, Stolfi C, Caruso R, Fina D, Sica G, Biancone L, Pallone F, Monteleone G (2008) Transcriptional and post-translational regulation of Flip, an inhibitor of Fas-mediated apoptosis, in human gut inflammation. Gut 57:1674–1680
Yerbes R, Palacios C, Reginato MJ, Lopez-Rivas A (2011) Cellular FLIP(L) plays a survival role and regulates morphogenesis in breast epithelial cells. Biochim Biophys Acta 1813:168–178
Acknowledgements
The authors wish to thank the skilful technical assistance from the technicians Birgit Deibjerg Kristensen, Anni Petersen, Hanne Fuglsang, Nine Scherling, Anne Hallander and Vibeke Voxen. This study was supported by grants from Fonden til Lægevidenskabens Fremme (the A.P. Møller Foundation), the Augustinus Foundation, and Aase and Ejnar Danielsens Foundation. JS holds a grant from the Danish Council for Independent Research. None of these research foundations had any influence on the outcome of this study or the interpretation of the results.
Disclosures
The authors have declared that no competing interests exist.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Seidelin, J.B., Coskun, M., Vainer, B. et al. ERK controls epithelial cell death receptor signalling and cellular FLICE-like inhibitory protein (c-FLIP) in ulcerative colitis. J Mol Med 91, 839–849 (2013). https://doi.org/10.1007/s00109-013-1003-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00109-013-1003-7