Skip to main content
NCBI home page
Journal of Cancer Research and Clinical Oncology logoLink to Journal of Cancer Research and Clinical Oncology
. 1997 Feb;123(2):100–106. doi: 10.1007/BF01269887

Unstimulated human acute myelogenous leukemia blasts secrete matrix metalloproteinases

Akinobu Matsuzaki 1, Anna Janowska-Wieczorek 1,✉
PMCID: PMC12201833  PMID: 9030248

Abstract

Purpose: The secretion of metalloproteinases was examined, especially the 92-kDa and 72-kDa type IV collagenases/gelatinases, and their role in the degradation of reconstituted basement membrane (Matrigel) by leukemic blasts.Methods and results: Leukemic blasts were obtained from the peripheral blood of 11 patients diagnosed with acute myelogenous leukemia (AML). After incubation of the AML blasts in serumfree cultures, conditioned media were collected and examined by zymography. The 92-kDa gelatinase was detected in ten AML patients after 2 h and 24 h of incubation, and in five samples its activated form (83 kDa) was observed. 72-kDa gelatinase was detected in cell-conditioned media from four patients after 2 h and in media from ten patients after 24 h. Its activated forms (64–60 kDa) were observed in one of four samples after 2 h and in five of ten after 24 h. By contrast, normal peripheral mononuclear cells from healthy donors secreted only 92-kDa gelatinase after 24 h; the 72 kDa enzyme was not detectable. A specific inhibitor of metalloproteinases, 1, 10-phenanthroline, significantly reduced the in vitro invasion of AML blasts in a Matrigel assay and completely inhibited gelatinolytic activity in zymography.Conclusions: We concluded that primary, unstimulated peripheral-blood AML blasts secrete metalloproteinases, which may contribute to the in vitro degradation of reconstituted basement membrane.

Key Words: AML, In vitro invasion, Type IV collagenases/gelatinases

Abbreviations

AML

acute myelogenous leukemia

MMP

matrix-degrading metalloproteinase

MNC

mononuclear cells

IMDM

Iscove's modified Dulbecco's medium

BSA

bovine serum albumin

References

  1. Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN (1987) A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47:3239–3245 [PubMed] [Google Scholar]
  2. Albini A, Melchiori A, Santi L, Liotta A, Brown PD, Stetler-Stevenson WG (1991) Tumor cell invasion inhibited by TIMP-2. J Natl Cancer Inst 83:775–779 [DOI] [PubMed] [Google Scholar]
  3. Birkedal-Hansen H, Moore WGI, Bodden MK, Windsor LJ, Birkedal-Hansen B, DeCarlo A, Engler JA (1993) Matrix metalloproteinases: a review. Crit Rev Oral Biol Med 4:197–250 [DOI] [PubMed] [Google Scholar]
  4. Borregaard N, Sehested M, Nielsen BS, Sebgelov H, Kjeldsen L (1995) Biosynthesis of granule proteins in normal human bone marrow cells. Gelatinase is a marker of terminal neutrophil differentiation. Blood 85:812–817 [PubMed] [Google Scholar]
  5. Branch DR, Shah A, Guilbert LJ (1991) A specific and reliable bioassay for the detection of femtomolar levels of human and murine turmor necrosis factors. J Immunol Methods 143: 251–261 [DOI] [PubMed] [Google Scholar]
  6. Campbell EJ, Cury JD, Shapiro SD, Goldberg GI, Welgus HG (1991) Neutral proteinases of human mononuclear phagocytes: cellular differentiation markedly alters cell phenotypes for serine proteinases, metalloproteinases and tissue inhibitor of metalloproteinases. J Immunol 146:1286–1293 [PubMed] [Google Scholar]
  7. Campo E, Merino MJ, Tavassoli FA, Charonis AS, Stetler-Stevenson WG, Liotta LA (1992) Evaluation of basement membrane components and the 72kDa type IV collagenase in serous tumors of the ovary. Am J Surg Pathol 16:500–507 [DOI] [PubMed] [Google Scholar]
  8. Cockett MI, Birch ML, Murphy G, Hart I, Docherty PJ (1994) Metalloproteinase domain structure, cellular invasion and metastasis. Biochem Soc Trans 22:55–57 [DOI] [PubMed] [Google Scholar]
  9. Davies B, Brown PD, East N, Crimmin MJ, Balkwill FR (1993) A synthetic matrix metalloproteinase inhibitor decreases tumor burden and prolongs survival of mice bearing human ovarian carcinoma xenografts. Cancer Res 53:2087–2091 [PubMed] [Google Scholar]
  10. DeClerck YA, Yean TD, Chan D, Shimoda H, Langley KE (1991) Inhibition of tumor invasion of smooth muscle cell layers by recombinant human metalloproteinase inhibitor. Cancer Res 51:2151–2157 [PubMed] [Google Scholar]
  11. D'Errico A, Garbisa S, Liotta LA, Castronovo V, Stetler-Stevenson WG, Grigioni WF (1991) Augmentation of type IV collagenase, laminin receptor, and Ki67 proliferation antigen associated with human colon, gastric, and breast carcinoma progression. Mod Pathol 4:239–246 [PubMed] [Google Scholar]
  12. Freireich EJ (1984) Acute leukemia: a prototype of disseminated cancer. Cancer 53:2026–2032 [DOI] [PubMed] [Google Scholar]
  13. Fridman R, Toth M, Pena D, Mobashery S (1995) Activation of progelatinase B (MMP-9) by gelatinase A (MMP-2). Cancer Res 55:2548–2555 [PubMed] [Google Scholar]
  14. Garbisa S, Pozzati R, Muschel RJ, Saffiotti U, Ballin M, Goldfarb RH, Khoury G, Liotta LA (1987) Secretion of type IV collagenolytic protease and metastatic phenotype: induction by transfection with c-Ha-ras burt not c-Has-ras plus Ad2-Ela. Cancer Res 47:1523–1528 [PubMed] [Google Scholar]
  15. Garbisa S, Scagliotti G, Masiero L, Di Francesco C, Caenazzo C, Onisto M, Micela M, Stetler-Stevenson WG, Liotta LA (1992) Correlation of serum metalloproteinase levels with lung cancer metastasis and response to therapy. Cancer Res 52: 4548–4549 [PubMed] [Google Scholar]
  16. Graubert T, Johnston J, Berliner N (1993) Cloning and expression of the cDNA encoding mouse neutrophil gelatinase: demonstration of coordinate secondary granule protein gene expression during terminal neutrophil maturation. Blood 82:3192–3197 [PubMed] [Google Scholar]
  17. Heussen C, Dowdle EB (1980) Electrophoretic analysis of plasminogen activators in polyacrylamide gel containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 102: 196–202 [DOI] [PubMed] [Google Scholar]
  18. Janiak M, Hashmi HR, Janowska-Wieczorek A (1994) Use of the Matrigel-based assay to measure the invasiveness of leukemic cells. Exp Hematol 22:559–565 [PubMed] [Google Scholar]
  19. Janowska-Wieczorek A, Kossakowska AE, Belch AR, Edwards DR (1993) Transcripts for tissue inhibitors of metalloproteinases (TIMPs) but not for collagenases are found in leukemic blasts (abstract). Proc Am Assoc Cancer Res 34:423a [Google Scholar]
  20. Janowska-Wieczorek A, Hashmi HR, Sawicki G (1995) Matrix-degrading enzymes expressed on the surface of leukemic cells and their role in the degradation of basement membrane (abstract). Blood 10 [Suppl 1]:47a [Google Scholar]
  21. Kane SE, Gottesman MM (1990) The role of cathepsin L in malignant transformation. Semin Cancer Biol 1:127–136 [PubMed] [Google Scholar]
  22. Kjeldsen L, Bjerrum OW, Hovgaard D, Johnsen AH, Sehested M, Borregaard N (1992) Human neutrophil gelatinase: a marker for circulating blood neutrophils. Purification and quantitation by enzyme-linked immunosorbent assay Eur J Haematol 49: 180–191 [DOI] [PubMed] [Google Scholar]
  23. Kobayashi M, Hamada J, Li YQ, Shinobu N, Imamura M, Okada F, Takeichi N, Hosokawa M (1995) A possible role of 92kDa type IV collagenases in the extramedullary tumor formation in leukemia. JpJ Cancer Res 86:298–303 [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kohn EC, Liotta LA (1995) Molecular insights into cancer invasion: strategies for prevention and intervention. Cancer Res 55:1856–1862 [PubMed] [Google Scholar]
  25. Leppert D, Waubant E, Galardy R, Bunnett NW, Hauser SL (1995) T cell gelatinases mediate basement membrane transmigration in vitro. J Immunol 154:4379–4389 [PubMed] [Google Scholar]
  26. Liotta LA, Tryggvason K, Garbisa S, Hart I, Foltz CM, Shafie S (1980) Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature 284:67–68 [DOI] [PubMed] [Google Scholar]
  27. Masure S, Proost P, van Damme J, Opdenakker G (1991) Purification and identification of 91-kDa neutrophil gelatinase. Release by the activating peptide interleukin-8. Eur J Biochem 198:391–398 [DOI] [PubMed] [Google Scholar]
  28. Montgomery AMP, Sabzevari H, Reisfeld A (1993) Production and regulation of gelatinase B by human T-cells. Biochim Biophys Acta 1176:265–268 [DOI] [PubMed] [Google Scholar]
  29. Morodomi T, Suzuki K, Sasaguri Y, Morimatsu M, Nagase H (1992) Comparative studies of a 92-kDa gelatinolytic metalloproteinase from U937 cells and matrix metalloproteinase 2 from human rheumatoid synovial fibroblasts. Matrix Suppl 1:87–88 [PubMed] [Google Scholar]
  30. Murphy G, Reynolds JL (1993) Extracellular matrix degradation. In: Royce PM, Steinmann B (eds) Connective tissue and its heritable disorders. Wiley-Liss, New York, pp 287–316 [Google Scholar]
  31. Pluznik DH, Fridman R, Reich R (1992) Correlation in the expression of tpe IV collagenases and the invasive and chemotactic abilities of myelomonocytic cells during differentiation into macrophages. Exp Hematol 20:57–63 [PubMed] [Google Scholar]
  32. Pyke C, Ralfkiaer E, Huhtala P, Hurskainen T, Dano K, Tryggvason K (1992) Localization of messenger RNA for M(r) 72,000 and 92,000 type IV collagenases in human skin cancers by in situ hybridization. Cancer Res 52:1336–1341 [PubMed] [Google Scholar]
  33. Ries C, Kolb H, Petrides PE (1994) Regulation of 92-kDa gelatinase in HL-60 leukemia cells: tumor necrosis factor-α as an autocrine stimulus for basal- and phorbol ester-induced secretion. Blood 12:3638–3646 [PubMed] [Google Scholar]
  34. Sato H, Takino T, Okada Y, Cao J, Shinagawa A, Yamamoto E, Seiki M (1994) A matrix metalloproteinase expressed on the surface of invasive tumor cells. Nature 370:61–65 [DOI] [PubMed] [Google Scholar]
  35. Schultz RM, Siberman S, Persky B, Bajlowski AS, Carmichael DF (1988) Inhibition by human recombinant tissue inhibitor of metalloproteinases of human amnion invasion and lung colonization by murine B-16F10 melanoma cells. Cancer Res 48:5539–5545 [PubMed] [Google Scholar]
  36. Sledge GW, Qulali M, Goulet R, Bone EA, Fife R (1995) Effect of matrix metalloproteinases inhibitor Batimastat on breast cancer regrowth and metastasis in athymic mice. J Natl Cancer Institute 87:1546–50 [DOI] [PubMed] [Google Scholar]
  37. Stetler-Stevenson WG (1994) Progelatinase A activation during tumor cell invasion. Invasion Metastasis 14:259–268 [PubMed] [Google Scholar]
  38. Stetler-Stevenson WG, Aznavoorian S, Liotta LA (1993) Tumor cell interactions with the extracellular matrix during invasion and metastasis. Annu Rev Cell Biol 9:541–573 [DOI] [PubMed] [Google Scholar]
  39. Tryggvason K, Hoyhtya M, Pyke C (1993) Type IV collagenases in invasive tumors. Breast Cancer Res Treat 24:209–218 [DOI] [PubMed] [Google Scholar]
  40. Tschesche H, Kaeuper V, Kraemer S, Michaelis J, Oberhoff R, Reinke H (1992) Latent collagenases and gelatinases from human neutrophils and their activation. Matrix Suppl 1:245–255 [PubMed] [Google Scholar]
  41. Wang X, Fu X, Brown PD, Crimmin MJ, Hoffman RM (1994) Matrix metalloproteinase inhibitor BB-94 (Batimastat) inhibits human colon tumor growth and spread in a patientlike orthotopic model in nude mice. Cancer Res 54:4726–4728 [PubMed] [Google Scholar]
  42. Watanabe H, Nakanishi I, Yamashita K, Hayakawa T, Okada Y (1993) Matrix metalloproteinase-9 (92 kDa gelatinase/type IV collagenase) from U937 monoblastoid cells: correlation with cellular invasion. J Cell Sci 104:991–999 [DOI] [PubMed] [Google Scholar]
  43. Welgus HG, Senior RM, Parks WC, Kahn AJ, Ley TJ, Shapiro SD, Campbell EJ (1992) Neutral proteinase expression by human mononuclear phagocytes: a prominent role of cellular differentiation. Matrix Suppl 1:363–367 [PubMed] [Google Scholar]
  44. Yamamoto M, Mohanam S, Sawaya R, Fuller GN, Seiki M, Sato H, Gokaslan ZL, Liotta LA, Nicolson GL, Rao JS (1996) Differential expression of membrane-type matrix metalloproteinases and its correlation with gelatinase A activation in human malignant brain tumors in vivo and in vitro. Cancer Res 56:384–392 [PubMed] [Google Scholar]
  45. Zucker S, Lysik RM, Zarrabi MH, Moll U (1993) M(r) 92,000 type IV collagenase is increased in plasma of patients with colon cancer and breast cancer. Cancer Res 53:140–146 [PubMed] [Google Scholar]

Articles from Journal of Cancer Research and Clinical Oncology are provided here courtesy of Springer

RESOURCES