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Pathology & Oncology Research

, Volume 7, Issue 4, pp 251–259 | Cite as

Expression of thrombospondin-1 in human pancreatic adenocarcinomas: Role in matrix metalloproteinase-9 production

  • Xiaohua Qian
  • Vicki L. Rothman
  • Roberto F. Nicosia
  • George P. Tuszynski
Article

Abstract

Human pancreatic adenocarcinoma, an aggressive malignant disease, shows a strong desmoplastic reaction characterized by a remarkable proliferation of interstitial connective tissues. Thrombospondin-1 (TSP-1), a 450 kDa platelet and matrix glycoprotein, has been implicated in tumor invasion, angiogenesis and metastasis. TSP-1 and MMP-9 expression in pancreatic adenocarcinoma and control pancreas tissues was measured by immunohistochemistry. TSP-1 expression in pancreatic carcinoma cell lines, fibroblasts, and endothelial cells was measured by a competitive TSP-1 enzyme linked immunosorbent assay (ELISA). The effect of TSP-1 on MMP-9 production in pancreatic carcinoma cell lines was measured by zymography and Western blot analysis. Eighty five per cent (23/27) of cases of pancreatic adenocarcinoma showed increased TSP-1 staining in the desmoplastic stroma adjacent to tumor cells. No specific positive staining for TSP-1 was observed in the normal pancreatic tissues and the inflammatory areas. TSP-1 localized in tumor stroma surrounding the tumor cells expressing MMP-9. Using TSP-1 competitive ELISA, the secretion of TSP-1 by different pancreatic cancer cell lines into culture medium varied from 11.45 ±14.08 to 275.82 ±45.56 ng/106 cells/24 hours. The amounts of TSP-1 detected in both culture media and cell extracts from fibroblasts or endothelial cells were at least 2–3 fold higher than those from pancreatic cancer cells. TSP-1 augmented the production of matrix metalloproteinase-9, a matrix degrading enzyme, in pancreatic cancer cellsin vitro. Stromally-derived TSP-1 up-regulates the production of MMP-9 by pancreatic adenocarcinoma. These data are consistent with the conclusion that TSP-1-rich stroma is involved in regulating matrix remodeling in tumor invasion.

Keywords

extracellular matrix thrombospondin-1 matrix metalloproteinase pancreatic cancer tumor invasion tissue inhibitor of metalloproteinase 

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References

  1. 1.2
    Parker SL, Tong T, Bolden S, Wingo PA: Cancer Statistics, 1996. CA-A Cancer J Clin 65:5–27, 1996.CrossRefGoogle Scholar
  2. 2.2
    Almoguera C, Shibata D, Forrester K: Most human carcinomas of the human exocrine pancreas contain mutant of c-K-ras genes. Cell 53:549–554, 1988.PubMedCrossRefGoogle Scholar
  3. 3.2
    Brentnall TA, Chen R, Lee JG, et al: Microsatellite instability and K-ras mutations associated with pancreatic adenocarcinoma and pancreatitis. Cancer Res 55:4264–4267, 1995.PubMedGoogle Scholar
  4. 4.2
    Hohne MW, Halatsch M, Kahl GF, et al: Frequent loss of expression of the potential tumor suppressor gene DCC in ductal pancreatic adenocarcinoma. Cancer Res 52:2616–2619, 1992.PubMedGoogle Scholar
  5. 5.2
    Ruggeri BA, Huang LY, Berger D, et al: Molecular pathology of primary and metastatic ductal pancreatic lesions. Analyses of mutations and expression of the p53, mdm-2, and p21/WAF-1 genes in sporadic and familial lesions. Cancer 79:700–716, 1997.PubMedCrossRefGoogle Scholar
  6. 6.2
    Hahn SA, Schutte M, Hoque ATMS, et al:DPC4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science 271:350–353, 1996.PubMedCrossRefGoogle Scholar
  7. 7.2
    Hall PA, Lemoine NR: Models of Pancreatic Cancer. Can Surveys 16:135–155, 1993.Google Scholar
  8. 8.2
    Stetler-Stevenson WG, Aznavoorian S, Liotta LA: Tumor cell interactions with the extracellular matrix during invasion and metastasis. Annu Rev Cell Biol 9:541–573, 1993.PubMedCrossRefGoogle Scholar
  9. 9.2
    Tuszynski GP, Nicosia RF: Localization of thrombospondin and its cysteine-serine-valinethreonine-cysteine-glycine-specific receptor in human breast carcinoma. Lab Invest 70:228–233, 1994.PubMedGoogle Scholar
  10. 10.2
    Bertin N, Clezardin P, Kubiak R, et al: Thrombospondin-1 and -2messenger RNA expression in normal, benign, and neoplastic human breast tissues: correlation with prognostic factors, tumor angiogenesis, and fibroblastic desmoplasia. Cancer Res 57:596–399, 1997.Google Scholar
  11. 11.2
    Qian XH, Tuszynski GP: Expression of thrombospondin-1 in cancer: A role in tumor progression. Proc Soc Exp Biol Med 212:199–207, 1996.PubMedGoogle Scholar
  12. 12.2
    Roberts DD: Regulation of tumor growth and metastasis by thrombospondin-1; FASEB J 10:1183–1191, 1996.PubMedGoogle Scholar
  13. 13.2
    Wang TN, Qian XH, Granick MS, et al: Thrombospondin-1 (TSP-1) promotes the invasive properties of human breast cancer. J Surg Res 63:39–43, 1996.PubMedCrossRefGoogle Scholar
  14. 14.2
    Qian X, Wang TN, Rothman VL, et al: Thrombospondin-1 modulates angiogenesisin vitro by up-regulation of matrix metalloproteinase-9 in endothelial cells. Exp Cell Res 235:403–412, 1997.PubMedCrossRefGoogle Scholar
  15. 15.2
    Albo D, Rothman VL, Roberts DD, et al: Tumor cell thrombospondin-1 regulates tumor cell adhesion and invasion through the urokinase plasminogen activator receptor. Br J Cancer 83:298–306, 2000.PubMedCrossRefGoogle Scholar
  16. 16.2
    Ray JM, Stetler-Stevenson WG: The role of matrix metelloproteases and their inhibitors in tumor invasion, metastasis and angiogenesis. Europ Resp J 7:2062–2072, 1994.Google Scholar
  17. 17.2
    Murphy G, Crabbe T: Gelatinases A and B. Methods Enzymol 248:470–484, 1995.PubMedCrossRefGoogle Scholar
  18. 18.2
    Okada Y, Gonoji Y, Naka K, et al: Matrix metalloproteinase 9 (92-kDa Gelatinase/type IV collagenase) from HT 1080 human fibrosarcoma cells; J Biol Chem 267:21712–21719, 1992.PubMedGoogle Scholar
  19. 19.2
    Hua J, Muschel RJ: Inhibition of matrix metalloproteinase 9 expression by a ribozyme blocks metastasis in a rat sarcoma model system. Cancer Res 56:5279–5284, 1996.PubMedGoogle Scholar
  20. 20.2
    Lengyel E, Gum R, Juarez J, et al: Induction of Mr 92,000 type IV collagenase expression in a squamous cell carcinoma cell line by fibroblasts. Cancer Res 55:963–967, 1995.PubMedGoogle Scholar
  21. 21.2
    Segain JP, Harb J, Grégoire M, et al: Induction of fibroblast gelatinase B expression by direct contact with cell lines derived from primary tumor but not from metastases. Cancer Res 56:5506–5512, 1996.PubMedGoogle Scholar
  22. 22.2
    Tuszynski GP, Switalska HI, Knudsen K: Methods of studying platelet-secreted proteins and the platelet cytoskeleton. Modern Methods Pharmacol 4:267–286, 1987.Google Scholar
  23. 23.2
    Tuszynski GP, Srivastava S, Switalska HI, et al: The interaction of human platelet thrombospondin with fibrinogen. J Biol Chem 260:12240–12245, 1985.PubMedGoogle Scholar
  24. 24.2
    Murphy-Ullrich JE, Schultz-Cherry S, Hook M; Transforming growth factor-b complexes with thrombospondin. Mol Biol Cell 3:181–188, 1992.PubMedGoogle Scholar
  25. 25.2
    Kinoshita T, Sato H, Takino T, et al: Processing of a precursor of 72-kilodalton type IV collagenase/gelatinase A by a recombinant membrane-type 1 matrix metalloproteinase. Cancer Res 56:2535–2538, 1996.PubMedGoogle Scholar
  26. 26.2
    Kleiner DE, Stetler-Stevenson WG: Quantitative zymography: Detection of Picogram quantities of gelatinases. Anal Biochem 218:325–329, 1994.PubMedCrossRefGoogle Scholar
  27. 27.2
    Himelstein BP, Canete-Soler R, Bernhard EJ, et al: Metalloproteinases in tumor progression: the contribution of MMP-9. Invas Metast 14:246–258, 1994.Google Scholar
  28. 28.2
    Vischer P, Beeck H, Voss B: Synthesis, intracellular processing and secretion of thrombospondin in human endothelial cells. Eur J Biochem 153:435–443, 1985.PubMedCrossRefGoogle Scholar
  29. 29.2
    Clezardin P: Expression of thrombospondin by cells in culture. In: Lahav J, ed. Thrombospondin. Boca Raton: CRC Press, 41–61, 1993.Google Scholar
  30. 30.2
    Goldberg GI, Strongin A, Collier IE, et al: Interaction of 92-kDa type IV collagenase with the tissue inhibitor of metalloproteinases prevents dimerization, complex formation with interstitial collagenase, and activation of the proenzyme with stromelysin; J Biol Chem 267:4583–4591, 1992.PubMedGoogle Scholar
  31. 31.2
    Lyons-Giordano B, Brinker JM, Kefalides NA: Heparin increases mRNA levels of thrombospondin but not fibronectin in human vascular smooth muscle cells; Biochem Biophys Res Comm 162:1100–1104, 1989.PubMedCrossRefGoogle Scholar
  32. 32.2
    Clezardin P, Frappart L, Clerget M, et al: Expression of thrombospondin (TSP1) and its receptors (CD36 and CD51) in normal, hyperplastic, and neoplastic human breast; Cancer Res 53:1421–1430, 1993.PubMedGoogle Scholar
  33. 33.2
    Friess H, Yamanaka Y, Büchler M, et al: Enhanced expession of transforming growth factor b isoforms in pancreatic cancer correlates with decreased survival. Gastroenterology 105:1846–856, 1993.PubMedGoogle Scholar
  34. 34.2
    Tikhonenko AT, Black DJ, Linial ML: Viral Myc oncoproteins in infected fibroblasts down-modulate thrombospondin-1, a possible tumor suppressor gene; J Biol Chem 271:30741–30747, 1996.PubMedCrossRefGoogle Scholar
  35. 35.2
    John AS, Tuszynski GP: The Role of Matrix Metalloproteinases in Tumor Angiogenesis and Tumor Metastasis. Pathol Oncol Res 7:14–23, 2001.PubMedCrossRefGoogle Scholar
  36. 36.2
    Murphy G, Willenbrock F: Tissue inhibitors of matrix metalloendopeptidases. Methods Enzymol 248:496–510, 1995.PubMedCrossRefGoogle Scholar
  37. 37.2
    Gress TM, Müller-Pillasch F, Lerch MM, et al: Expression andin-situ localization of genes coding for extracellular matrix proteins and extracellular matrix degrading proteases in pancreatic cancer. Int J Cancer 62:407–413, 1995.PubMedCrossRefGoogle Scholar
  38. 38.2
    Hirata M, Itoh M, Tsuchida A, et al: Cholecystokinin receptor antagonist, loxiglumide, inhibits invasiveness of human pancreatic cancer cell lines. FEBS Lett 383:241–244, 1996.PubMedCrossRefGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2001

Authors and Affiliations

  • Xiaohua Qian
    • 2
  • Vicki L. Rothman
    • 1
  • Roberto F. Nicosia
    • 2
    • 3
  • George P. Tuszynski
    • 1
    • 2
  1. 1.Department of SurgeryMCP Hahnemann UniversityPhiladelphiaUSA
  2. 2.Department of PathologyMCP Hahnemann UniversityPhiladelphiaUSA
  3. 3.Hahnemann UniversityPhiladelphiaUSA

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