Abstract
Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine affecting diverse cellular responses. TNF-α is cytotoxic in many systems, but it can also act as an anti-apoptotic signal to promote cell survival pathways activated through integrins and extracellular matrix components. This is particularly evident in cancer cells. To unravel the basis of resistance to TNF-α-induced apoptosis, human osteosarcoma MG-63 cell line was used. Our data showed that resistance to apoptosis was accompanied by high levels of TIMP-1 expression in part mediated by NF-κB activation, whereas under apoptotic conditions, in the presence of cycloheximide (CHX), TIMP-1 and αvβ3 integrin protein levels were significantly reduced. Silencing TIMP-1 using siRNA led to increased apoptosis following treatment with TNF-α, whereas exogenously-added recombinant TIMP-1 reduced the extent of apoptosis. Immunoprecipitation and confocal microscopy experiments demonstrated that TIMP-1 interacted with αvβ3 integrins. The biological role of this interaction was revealed by the use of echistatin, an antagonist of αvβ3 integrin. In the presence of echistatin, decreased protection against apoptosis by recombinant TIMP-1 was observed.
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References
Bieler G et al (2007) Distinctive role of integrin-mediated adhesion in TNF-induced PKB/Akt and NF-kappaB activation and endothelial cell survival. Oncogene 26:5722–5732
Boulday G et al (2004) Exogenous tissue inhibitor of metalloproteinase-1 promotes endothelial cell survival through activation of the phosphatidylinositol 3-kinase/Akt pathway. Ann NY Acad Sci 1030:28–36
Cheong JW et al (2003) Induction of apoptosis by apicidin, a histone deacetylase inhibitor, via the activation of mitochondria-dependent caspase cascades in human Bcr-Abl-positive leukemia cells. Clin Cancer Res 9:5018–5027
Chua CC et al (2002) TGF-beta1 inhibits multiple caspases induced by TNF-alpha in murine osteoblastic MC3T3-E1 cells. Biochim Biophys Acta 1593:1–8
Fos C et al (2008) ICOS ligation recruits the p50alpha PI3K regulatory subunit to the immunological synapse. J Immunol 181:1969–1977
Gilbert L et al (2000) Inhibition of osteoblast differentiation by tumor necrosis factor-alpha. Endocrinology 141:3956–3964
Guedez L et al (1998) In vitro suppression of programmed cell death of B cells by tissue inhibitor of metalloproteinases-1. J Clin Invest 102:2002–2010
Inoue M et al (2000) Tumor necrosis factor alpha regulates alpha(v)beta5 integrin expression by osteoclast precursors in vitro and in vivo. Endocrinology 141:284–290
Jung KK et al (2006) Identification of CD63 as a tissue inhibitor of metalloproteinase-1 interacting cell surface protein. EMBO J 25:3934–3942
Karadag A, Fisher LW (2006) Bone sialoprotein enhances migration of bone marrow stromal cells through matrices by bridging MMP-2 to alpha(v)beta3-integrin. J Bone Miner Res 21:1627–1636
Karin M, Lin A (2002) NF-kappaB at the crossroads of life and death. Nat Immunol 3:221–227
Lazebnik YA et al (1994) Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371:346–347
Liu XW et al (2003) Tissue inhibitor of metalloproteinase-1 protects human breast epithelial cells against intrinsic apoptotic cell death via the focal adhesion kinase/phosphatidylinositol 3-kinase and MAPK signaling pathway. J Biol Chem 278:40364–40372
Locklin RM et al (2007) Selective targeting of death receptor 5 circumvents resistance of MG-63 osteosarcoma cells to TRAIL-induced apoptosis. Mol Cancer Ther 6:3219–3228
Matsumoto M et al (2000) Involvement of p38 mitogen-activated protein kinase signaling pathway in osteoclastogenesis mediated by receptor activator of NF-kappa B ligand (RANKL). J Biol Chem 275:31155–31161
Matter ML, Ruoslahti E (2001) A signaling pathway from the alpha5beta1 and alpha(v)beta3 integrins that elevates bcl-2 transcription. J Biol Chem 276:27757–27763
Ratnikov BI et al (2002) An alternative processing of integrin alpha(v) subunit in tumor cells by membrane type-1 matrix metalloproteinase. J Biol Chem 277:7377–7385
Retta SF et al (2001) Cross talk between beta(1) and alpha(V) integrins: beta(1) affects beta(3) mRNA stability. Mol Biol Cell 12:3126–3138
Sahni A, Francis CW (2004) Stimulation of endothelial cell proliferation by FGF-2 in the presence of fibrinogen requires alphavbeta3. Blood 104:3635–3641
Schrohl AS et al (2006) Primary tumor levels of tissue inhibitor of metalloproteinases-1 are predictive of resistance to chemotherapy in patients with metastatic breast cancer. Clin Cancer Res 12:7054–7058
Sun J et al (2007) Induction of apoptosis in osteogenic sarcoma cells by combination of tumor necrosis factor-related apoptosis inducing ligand and chemotherapeutic agents. Chin Med J (Engl) 120:400–404
Talamagas AA et al (2007) Abeta(1-40)-induced secretion of matrix metalloproteinase-9 results in sAPPalpha release by association with cell surface APP. Neurobiol Dis 28:304–315
Tsuboi M et al (1999) Tumor necrosis factor-alpha and interleukin-1beta increase the Fas-mediated apoptosis of human osteoblasts. J Lab Clin Med 134:222–231
Tzinia AK et al (2002) Effects of collagen IV on neuroblastoma cell matrix-related functions. Exp Cell Res 274:169–177
Uchida M et al (2000) Regulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by bone resorptive factors in osteoblastic cells. J Cell Physiol 185:207–214
Weise S et al (2008) Tissue inhibitor of metalloproteinase-1 mRNA production and protein secretion are induced by interleukin-1 beta in 3 T3-L1 adipocytes. J Endocrinol 198:169–174
Yoneda T, Hiraga T (2005) Crosstalk between cancer cells and bone microenvironment in bone metastasis. Biochem Biophys Res Commun 328:679–687
Zhivotovsky B (2003) Caspases: the enzymes of death. Essays Biochem 39:25–40
Zhou X et al (2004) Engagement of alphavbeta3 integrin regulates proliferation and apoptosis of hepatic stellate cells. J Biol Chem 279:23996–24006
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The authors thank Dr. Marina Sagnou for her expert assistance with confocal microscopy experiments and Drs. Dimitris Kletsas and Harris Pratsinis for their useful discussions.
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This research was funded by NCSR Demokritos, the General Secretariat for Research & Development (grant No 2005/ΣΕ01330081) and funds by BIOTREK, TRIAS and HALYPS companies.
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Tsagaraki, I., Tsilibary, E.C. & Tzinia, A.K. TIMP-1 interaction with αvβ3 integrin confers resistance to human osteosarcoma cell line MG-63 against TNF-α-induced apoptosis. Cell Tissue Res 342, 87–96 (2010). https://doi.org/10.1007/s00441-010-1025-1
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DOI: https://doi.org/10.1007/s00441-010-1025-1