Abstract
In the present work we investigated the role and biological significance of mitogen activated protein kinases (MAPK) in B-cell chronic lymphocytic leukaemia (B-CLL). The MAPK p38 was constitutively activated in B-CLL, but not in normal peripheral B cells. In addition, we demonstrated that the upstream kinases of p38, MKK3/6 were also constitutively activated in B-CLL cells. Furthermore, we determined by EMSA that the p38 MAP kinase pathway was not linked to the constitutive high expression of NF-κB, a critical survival factor of B-CLL cells. Recently, it has been shown that serum levels of angiogenic factors like VEGF, bFGF and MMP-9 are elevated in the serum of CLL patients and correlate with an unfavorable prognosis. We showed that the constitutive expression of MMP-9 was dependent on p38-activity and inhibition of p38 strongly downregulated MMP-9 expression. Coculture of B-CLL cells and stromal cells can protect spontaneous apoptosis of leukemic B cells. To determine the role of permanently activated p38 and MMP-9 expression, we cocultured B-CLL cells with bone marrow stromal cells. Survival of B-CLL cells on stroma was severely impaired when p38 was inhibited. Furthermore, blockade of MMP-9 activity also antagonized the antiapoptotic effect of stromal cells.
Similar content being viewed by others
References
Furman RR, Asgary Z, Mascarenhas JO, Liou HC, Schattner EJ . Modulation of NF-κB activity and apoptosis in chronic lymphocytic leukaemia. J Immunol 2000; 164: 2200–2206.
Frank DA, Mahajan S, Ritz J . B lymphocytes from patients with chronic lymphocytic leukaemia contain signal transducer, activator of transcription (STAT)1 and STAT3 constitutively phosphorylated on serine residues. J Clin Invest 1997; 100: 3140–3148.
Ringshausen I, Schneller F, Bogner C, Hipp S, Duyster J, Peschel C, Decker T . Constitutively activated PI 3-K is involved in the defect of apoptosis in B-CLL: association with PCK-δ. Blood 2002; 100: 3741–3748.
Whitmarsh AJ, Davis RJ . Transcritipion factor AP-1 regulation by mitogen-activated protein kinase signal transduction pathways. J Mol Med 1996; 74: 589–607.
Boulton TG, Nye SH, Robbins DJ, Radziejwska E, Morgenbesser SD, DePhino RA et al. ERKs: a family of protein-serine/threonine kinases that are activated, tyrosine-phosphorylated in response to insulin NGF. Cell 1991; 65: 663–675.
Barr RK, Bogoyevitch MA . The c-Jun N-terminal protein kinase familiy of mitogen-activated protein kinases. Int J Biochem Cell Biol 2001; 33: 1047–1063.
Lee JC, Laydon JT, McDonnell PC, Gallagher TF, Kumar S, Green D et al. A protein kinase involved in the regulation of inflammatory cytokine biosynthesis. Nature 1994; 372: 739–746.
Chang L, Karin M . Mammilian MAP kinase signaling cascades. Nature 2001; 410: 37–40.
Raingeaud J, Gupta S, Rogers JS, Dickens M, Han J, Ulevitch RJ et al. Pro-inflammatory cytokines, environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine, threonine. J Biol Chem 1995; 270: 7420–7426.
Han J, Lee JD, Jiang Y, Li Z, Feng L, Ulevitch RJ . Characterisation of the strucuture, function of a novel MAP kinase kinase (MKK6). J Biol Chem 1996; 271: 2886–2891.
Enslen H, Raingeaud J, Davis RJ . Selective activation of the p38 mitogen-activated protein (MAP) kinase isoforms by the MAP kinase kinase MKK3 and MKK6. J Biol Chem 1998; 273: 1741–1748.
Han J, Lee JD, Bibbs L, Ulevitch RJ . A MAP kinase targeted by endotoxin and hyperosmolarity in mammilian cells. Science 1994; 265: 808–811.
Lee JC, Laydon JT, McDonnell PC, Gallagher TF, Kumar S, Green D et al. A protein kinase involved in the regulation of inflammatory cytokine biosynthesis. Nature 1994; 372: 739–746.
Zhao M, New L, Kravchenko VV, Kato Y, Gram H, diPadova F et al. Regulation of the MEF2 family of transcription factors by p38. Mol Cell Biol 1999; 19: 21–30.
Craxton A, Shu G, Graves JD, Saklatvala J, Krebs EG, Clark EA . p38 MAPK is required for CD40-induced gene expression and proliferation in B lymphocytes. J Immunol 1998; 161: 3225–3236.
Tan Y, Rouse J, Zhang A, Cariati S, Cohen P, Comb MJ . FGF and stress regulate CREB and ATF-1 via a pathway involving p38 MAP kinase and MAPKAP kinase-2. EMBO J 1996; 15: 4629–4642.
Wang XZ, Ron D . Stress-induced phosphorylation and activation of the transcription factor CHOP by p38 MAP kinase. Science 1996; 272: 1347–1349.
Bauvois B, Dumont J, Mathiot C, Kolb JP . Production of MMP 9 in early stage B-CLL: supression by interferons. Leukemia 2002; 16: 791–798.
Molica S, Vitelli G, Levato D, Giannarelli D, Vacca A, Cuneo A et al. Increased serum levels of MMP-9 predict clinical outcome of patients with early B-CLL. Eur J Haematol 2003; 70: 373–378.
Esteve PO, Chicoine E, Robledo O, Aoudjit F, Descoteaux A, Potworowski EF et al. Protein kinase C-zeta regulates transcription of the matrix metalloproteinase-9 gene induced by IL-1 and TNF-alpha in glioma cells via NF-κ B. J Biol Chem 2002; 277: 35150–35155.
Huhtala P, Tuuttila A, Chow LT, Lohi J, Keski-Oja J, Tryggvason K . Complete structure of the human gene for 92-kDa type IV collagenase. Divergent regulation of expression for the 92- and 72-kilodalton enzyme genes in HT-1080 cells. J Biol Chem 1991; 266: 16485.
Simon C, Simon M, Vucelic G . The p38 SAPK pathway regulates the expression of the MMP-9 collagenase via AP-1-dependent promotor activation. Exp Cell Res 2001; 271: 344–355.
Mackman N, Brand K, Edington TS . Lipopolysaccharide-mediated transcriptional activation of the human tissue factor gene in THP-1 monocytic cells requires both activator protein 1 and nuclear factor kappa B binding sites. J Exp Med 1991; 174: 1517–1526.
Ward KW, Proksch JW, Azzarano LM, Mumaw JA, Roethke TJ, Stelman GJ et al. Preclinical pharmacokinetics of SB203580, a potent inhibitor of p38 MAPK. Xenobiotica 2001; 31: 783–797.
Lagneaux L, Delforge A, Bron D, DeBruyn C, Stryckmans P . Chronic lymphocytic leukemia B cells but not normal B cells are rescued from apoptosis by contact with normal bone marrow stromal cells. Blood 1998; 91: 2387–2396.
Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME . Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 1995; 270: 1326–1331.
Pepper C, Thomas A, Hoy T, Milligan D, Bentley P, Fegan C . The vitamin D3 analog EB1089 induces apoptosis via a p53-independent mechanism involving p38 MAP kinase activation and suppression of ERK activity in B-cell chronic lymphocytic leukemia cells in vitro. Blood 2003; 101: 2454–2460.
Pederson IM, Buhl AM, Klausen P, Geisler CH, Jurlander J . The chimeric anti-CD20 antibody rituximab induces apoptosis in B-cell chronic lymphoytic leukemia cells through a p38 mitogen activated protein-kinase-dependent-mechanism. Blood 2002; 99: 1314–1319.
Hideshima T, Akiyama M, Hayashi T, Richardson P, Schlossman R, Chauhan D et al. Targeting p38 MAP kinase inhibits multiple myeloma cell growth in the bone marrow milieu. Blood 2002; 101: 703–705.
Elenitoba-Johnson KS, Jenson SD, Abbott RT, Palais RA, Bohling SD, Lin Z et al. Involvement of multiple signaling pathways in follicular lymphoma transformation: p38-mitogen-activated protein kinase as a target for therapy. Proc Natl Acad Sci USA 2003; 100: 7259–7264.
Sthoeger ZM, Wakai M, Tse DB, Vinciguerra VP, Allen SL, Budman DR et al. Production of autoantibodies by CD-5 expressing B lymphocytes from patients with chronic lymphocytic leukemia. J Exp Med 1989; 169: 255.
Gearing AJ, Beckett P, Christodoulou M, Churchill M, Clements JM, Crimmin M et al. MMPs and processing of pro-TNF-alpha. J Leukoc Biol 1995; 57: 774–777.
Digel W, Stefanic M, Schoniger W, Buck C, Raghavachar A, Frickhofen N et al. Tumor necrosis factor induces proliferation of neoplastic B-cells from chronic lymphocytic leukemia. Blood 1989; 73: 1242–1246.
Petitclerc E, Stromblad S, von Schalscha TL, Mitjans F, Piulats J, Montgomery AM et al. Integrin aVb3 promotes M21 melanoma growth in human skin by regulating tumor cell survival. Cancer Res 1999; 59: 2724–2730.
Collins RJ, Verschurer LA, Harmon BV, Prentice RL, Pope JH, Kerr JF . Sponatneous programmed death (apoptosis) of B-chronic lymphocytic leukemia cells following their culture in vitro. Br J Haematol 1989; 71: 343–350.
Pederson IM, Kitada S, Leoni LM . Protection of CLL B cells by a follicular dentritic cell line is dependent on induction of Mcl-1. Blood 2002; 100: 1795–1801.
Burger JA, Burger M, Kipps TJ . CLL B cells express functional CXCR4 chemokine receptors that mediates spontaneous migration beneath bone marrow stromal cells. Blood 1999; 94: 3658–3667.
Rahman A, Anwar KN, Uddin S, Xu N, Ye RD, Platanias LC et al. Protein kinase C-δ regulates thrombin-induced ICAM-1 gene expression in endothelial cells via the activation of p38 mitogen-activated protein kinase. Mol Cell Biol 2001; 21: 5554–5565.
Huang J, Frischer JS, Serur A, Kadenhe A, Yokoi A, McCrudden KW et al. Regression of established tumors and metastases by potent vascular endothelial growth factor blockade. Proc Natl Acad Sci USA 2003; 100: 7785–7790.
Tanimura S, Asato K, Fujishiro SH, Kohno M . Specific blockade of the ERK pathway inhibits the invasiveness of tumor cells: down-regulation of matrix metalloproteinase-3/-9/-14 and CD44. Biochem Biophys Res Commun 2003; 304: 801–806.
Acknowledgements
This work was supported by a research grant from the Technical University of Munich (KKF 15-00) and a grant from the Deutsche Forschungsgemeinschaft DE 771.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ringshausen, I., Dechow, T., Schneller, F. et al. Constitutive activation of the MAPkinase p38 is critical for MMP-9 production and survival of B-CLL cells on bone marrow stromal cells. Leukemia 18, 1964–1970 (2004). https://doi.org/10.1038/sj.leu.2403544
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.leu.2403544
- Springer Nature Limited
Keywords
This article is cited by
-
The Akt signaling pathway determines the different proliferative capacity of chronic lymphocytic leukemia B-cells from patients with progressive and stable disease
Leukemia (2007)
-
Hyperforin inhibits MMP-9 secretion by B-CLL cells and microtubule formation by endothelial cells
Leukemia (2006)
-
High Mda-7 expression promotes malignant cell survival and p38 MAP kinase activation in chronic lymphocytic leukemia
Leukemia (2006)
-
VEGF-induced survival of chronic lymphocytic leukemia is independent of Bcl-2 phosphorylation
Leukemia (2005)