Matrix metalloproteinases in human melanoma cell lines and xenografts: increased expression of activated matrix metalloproteinase-2 (MMP-2) correlates with melanoma progression

Abstract

Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are involved in tumour progression and metastasis. In this study, we investigated the in vitro and in vivo expression patterns of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1 and TIMP-2 mRNA and protein in a previously described human melanoma xenograft model. This model consists of eight human melanoma cell lines with different metastatic behaviour after subcutaneous (s.c.) injection into nude mice. MMP-1 mRNA was detectable in all cell lines by reverse transcription polymerase chain reaction (RT-PCR), but the expression was too low to be detected by Northern blot analysis. No MMP-1 protein could be found using Western blotting. MMP-2 mRNA and protein were present in all cell lines, with the highest expression of both latent and active MMP-2 in the highest metastatic cell lines MV3 and BLM. MMP-3 mRNA was expressed in MV3 and BLM, and in the non-metastatic cell line 530, whereas MMP-3 protein was detectable only in MV3 and BLM. None of the melanoma cell lines expressed MMP-9. TIMP-1 and TIMP-2 mRNA and protein, finally, were present in all cell lines. A correlation between TIMP expression level and metastatic capacity of cell lines, however, was lacking. MMP and TIMP mRNA and protein expression levels were also studied in s.c. xenograft lesions derived from a selection of these cell lines. RT-PCR analysis revealed that MMP-1 mRNA was present in MV3 and BLM xenografts, and to a lesser extent in 530. Positive staining for MMP-1 protein was found in xenograft lesions derived from both low and high metastatic cell lines, indicating an in vivo up-regulation of MMP-1. MMP-2 mRNA was detectable only in xenografts derived from the highly metastatic cell lines 1F6m, MV3 and BLM. In agreement with the in vitro results, the highest levels of both latent and activated MMP-2 protein were observed in MV3 and BLM xenografts. With the exception of MMP-9 mRNA expression in 530 xenografts, MMP-3, MMP-9, and TIMP-1 mRNA and protein were not detectable in any xenograft, indicating a down-regulated expression of MMP-3 and TIMP-1 in vivo. TIMP-2 mRNA and protein were present in all xenografts; interestingly, the strongest immunoreactivity of tumour cells was found at the border of necrotic areas. Our study demonstrates that of all tested components of the matrix metalloproteinase system, only expression of activated MMP-2 correlates with increased malignancy in our melanoma xenograft model, corroborating an important role of MMP-2 in human melanoma invasion and metastasis.

References 40

1. 1.

   Ahonen M., Baker A. H., Kähäri V. M. Adenovirus-mediated gene delivery of tissue inhibitor of metalloproteinases-3 inhibits invasion and induces apoptosis in melanoma cells. Cancer Res. 1998;58(11):2310-2315.

   Not in our library

2. 2.

   Butler G. S., Butler M. J., Atkinson S. J., Will H., Tamura T., Schade van Westrum S., Crabbe T., Clements J., d'Ortho M. P., Murphy G. The TIMP2 membrane type 1 metalloproteinase "receptor" regulates the concentration and efficient activation of progelatinase A. A kinetic study. J Biol Chem. 1998;273(2):871-880.

   Not in our library10.1074/jbc.273.2.871

3. 3.

   Chambers A. F., Matrisian L. M. Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst. 1997;89(17):1260-1270.

   Not in our library10.1093/jnci/89.17.1260

4. 4.

   Coussens L. M., Werb Z. Matrix metalloproteinases and the development of cancer. Chem Biol. 1996;3(11):895-904.

   Not in our library10.1016/s1074-5521(96)90178-7

5. 5.

   Durko M., Navab R., Shibata H. R., Brodt P. Suppression of basement membrane type IV collagen degradation and cell invasion in human melanoma cells expressing an antisense RNA for MMP-1. Biochim Biophys Acta. 1997;1356(3):271-280.

   Not in our library10.1016/s0167-4889(97)00004-9

6. 6.

   Ferrier C. M., van Geloof W. L., de Witte H. H., Kramer M. D., Ruiter D. J., van Muijen G. N. Epitopes of components of the plasminogen activation system are re-exposed in formalin-fixed paraffin sections by different retrieval techniques. J Histochem Cytochem. 1998;46(4):469-476.

   Not in our library10.1177/002215549804600406

7. 7.

   Gomez D. E., Alonso D. F., Yoshiji H., Thorgeirsson U. P. Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. Eur J Cell Biol. 1997;74(2):111-122.

   Not in our library

8. 8.

   Grant G. M., Cobb J. K., Castillo B., Klebe R. J. Regulation of matrix metalloproteinases following cellular transformation. J Cell Physiol. 1996;167(1):177-183.

   Not in our library10.1002/(sici)1097-4652(199604)167:1<177::aid-jcp21>3.0.co;2-7

9. 9.

   Heppner K. J., Matrisian L. M., Jensen R. A., Rodgers W. H. Expression of most matrix metalloproteinase family members in breast cancer represents a tumor-induced host response. Am J Pathol. 1996;149(1):273-282.

   Not in our library

10. 10.

    Imren S., Kohn D. B., Shimada H., Blavier L., DeClerck Y. A. Overexpression of tissue inhibitor of metalloproteinases-2 retroviral-mediated gene transfer in vivo inhibits tumor growth and invasion. Cancer Res. 1996;56(13):2891-2895.

    Not in our library

11. 11.

    Kanayama H., Yokota K., Kurokawa Y., Murakami Y., Nishitani M., Kagawa S. Prognostic values of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression in bladder cancer. Cancer. 1998;82(7):1359-1366.

    Not in our library

12. 12.

    Khokha R. Suppression of the tumorigenic and metastatic abilities of murine B16-F10 melanoma cells in vivo by the overexpression of the tissue inhibitor of the metalloproteinases-1. J Natl Cancer Inst. 1994;86(4):299-304.

    Not in our library10.1093/jnci/86.4.299

13. 13.

    Khokha R., Zimmer M. J., Wilson S. M., Chambers A. F. Up-regulation of TIMP-1 expression in B16-F10 melanoma cells suppresses their metastatic ability in chick embryo. Clin Exp Metastasis. 1992;10(6):365-370.

    Not in our library10.1007/bf00133464

14. 14.

    Lampert K., Machein U., Machein M. R., Conca W., Peter H. H., Volk B. Expression of matrix metalloproteinases and their tissue inhibitors in human brain tumors. Am J Pathol. 1998;153(2):429-437.

    Not in our library10.1016/s0002-9440(10)65586-1

15. 15.

    MacDougall J. R., Bani M. R., Lin Y., Rak J., Kerbel R. S. The 92-kDa gelatinase B is expressed by advanced stage melanoma cells: suppression by somatic cell hybridization with early stage melanoma cells. Cancer Res. 1995;55(18):4174-4181.

    Not in our library

16. 16.

    MacDougall J. R., Matrisian L. M. Contributions of tumor and stromal matrix metalloproteinases to tumor progression, invasion and metastasis. Cancer Metastasis Rev. 1995;14(4):351-362.

    Not in our library10.1007/bf00690603

17. 17.

    Montgomery A. M., Mueller B. M., Reisfeld R. A., Taylor S. M., DeClerck Y. A. Effect of tissue inhibitor of the matrix metalloproteinases-2 expression on the growth and spontaneous metastasis of a human melanoma cell line. Cancer Res. 1994;54(20):5467-5473.

    Not in our library

18. 18.

    Mueller B. M. Different roles for plasminogen activators and metalloproteinases in melanoma metastasis. Curr Top Microbiol Immunol. 1996;213(Pt 1):65-80.

    Not in our library10.1007/978-3-642-61107-0_5

19. 19.

    Nagase H. Activation mechanisms of matrix metalloproteinases. Biol Chem. 1997;378(3-4):151-160.

    Not in our library

20. 20.

    Nakahara H., Howard L., Thompson E. W., Sato H., Seiki M., Yeh Y., Chen W. T. Transmembrane/cytoplasmic domain-mediated membrane type 1-matrix metalloprotease docking to invadopodia is required for cell invasion. Proc Natl Acad Sci U S A. 1997;94(15):7959-7964.

    Not in our library10.1073/pnas.94.15.7959

21. 21.

    Newell K. J., Witty J. P., Rodgers W. H., Matrisian L. M. Expression and localization of matrix-degrading metalloproteinases during colorectal tumorigenesis. Mol Carcinog. 1994;10(4):199-206.

    Not in our library10.1002/mc.2940100404

22. 22.

    Nielsen B. S., Timshel S., Kjeldsen L., Sehested M., Pyke C., Borregaard N., Danø K. 92 kDa type IV collagenase (MMP-9) is expressed in neutrophils and macrophages but not in malignant epithelial cells in human colon cancer. Int J Cancer. 1996;65(1):57-62.

    Not in our library10.1002/(sici)1097-0215(19960103)65:1<57::aid-ijc10>3.0.co;2-f

23. 23.

    Nomura H., Sato H., Seiki M., Mai M., Okada Y. Expression of membrane-type matrix metalloproteinase in human gastric carcinomas. Cancer Res. 1995;55(15):3263-3266.

    Not in our library

24. 24.

    Ray J. M., Stetler-Stevenson W. G. Gelatinase A activity directly modulates melanoma cell adhesion and spreading. EMBO J. 1995;14(5):908-917.

    Not in our library10.1002/j.1460-2075.1995.tb07072.x

25. 25.

    Sato H., Takino T., Okada Y., Cao J., Shinagawa A., Yamamoto E., Seiki M. A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature. 1994;370(6484):61-65.

    Not in our library10.1038/370061a0

26. 26.

    Sreenath T., Matrisian L. M., Stetler-Stevenson W., Gattoni-Celli S., Pozzatti R. O. Expression of matrix metalloproteinase genes in transformed rat cell lines of high and low metastatic potential. Cancer Res. 1992;52(18):4942-4947.

    Not in our library

27. 27.

    Stetler-Stevenson W. G., Aznavoorian S., Liotta L. A. Tumor cell interactions with the extracellular matrix during invasion and metastasis. Annu Rev Cell Biol. 1993;9:541-573.

    Not in our library10.1146/annurev.cb.09.110193.002545

28. 28.

    Strongin A. Y., Collier I., Bannikov G., Marmer B. L., Grant G. A., Goldberg G. I. Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease. J Biol Chem. 1995;270(10):5331-5338.

    Not in our library10.1074/jbc.270.10.5331

29. 29.

    Sugiura Y., Shimada H., Seeger R. C., Laug W. E., DeClerck Y. A. Matrix metalloproteinases-2 and -9 are expressed in human neuroblastoma: contribution of stromal cells to their production and correlation with metastasis. Cancer Res. 1998;58(10):2209-2216.

    Not in our library

30. 30.

    Tokuraku M., Sato H., Murakami S., Okada Y., Watanabe Y., Seiki M. Activation of the precursor of gelatinase A/72 kDa type IV collagenase/MMP-2 in lung carcinomas correlates with the expression of membrane-type matrix metalloproteinase (MT-MMP) and with lymph node metastasis. Int J Cancer. 1995;64(5):355-359.

    Not in our library10.1002/ijc.2910640513

31. 31.

    Valente P., Fassina G., Melchiori A., Masiello L., Cilli M., Vacca A., Onisto M., Santi L., Stetler-Stevenson W. G., Albini A. TIMP-2 over-expression reduces invasion and angiogenesis and protects B16F10 melanoma cells from apoptosis. Int J Cancer. 1998;75(2):246-253.

    Not in our library10.1002/(sici)1097-0215(19980119)75:2<246::aid-ijc13>3.0.co;2-b

32. 32.

    Van Muijen G. N., Cornelissen L. M., Jansen C. F., Figdor C. G., Johnson J. P., Bröcker E. B., Ruiter D. J. Antigen expression of metastasizing and non-metastasizing human melanoma cells xenografted into nude mice. Clin Exp Metastasis. 1991;9(3):259-272.

    Not in our library10.1007/bf01753729

33. 33.

    Väisänen A., Kallioinen M., Taskinen P. J., Turpeenniemi-Hujanen T. Prognostic value of MMP-2 immunoreactive protein (72 kD type IV collagenase) in primary skin melanoma. J Pathol. 1998;186(1):51-58.

    Not in our library10.1002/(sici)1096-9896(199809)186:1<51::aid-path131>3.0.co;2-p

34. 34.

    Väisänen A., Tuominen H., Kallioinen M., Turpeenniemi-Hujanen T. Matrix metalloproteinase-2 (72 kD type IV collagenase) expression occurs in the early stage of human melanocytic tumour progression and may have prognostic value. J Pathol. 1996;180(3):283-289.

    Not in our library10.1002/(sici)1096-9896(199611)180:3<283::aid-path662>3.0.co;2-3

35. 35.

    Westphal J. R., van't Hullenaar R. G., van der Laak J. A., Cornelissen I. M., Schalkwijk L. J., van Muijen G. N., Wesseling P., de Wilde P. C., Ruiter D. J., de Waal R. M. Vascular density in melanoma xenografts correlates with vascular permeability factor expression but not with metastatic potential. Br J Cancer. 1997;76(5):561-570.

    Not in our library10.1038/bjc.1997.427

36. 36.

    Woolley D. E., Grafton C. A. Collagenase immunolocalization studies of cutaneous secondary melanomas. Br J Cancer. 1980;42(2):260-265.

    Not in our library10.1038/bjc.1980.225

37. 37.

    Yamamoto M., Mohanam S., Sawaya R., Fuller G. N., Seiki M., Sato H., Gokaslan Z. L., Liotta L. A., Nicolson G. L., Rao J. S. Differential expression of membrane-type matrix metalloproteinase and its correlation with gelatinase A activation in human malignant brain tumors in vivo and in vitro. Cancer Res. 1996;56(2):384-392.

    Not in our library

38. 38.

    Zucker S., Drews M., Conner C., Foda H. D., DeClerck Y. A., Langley K. E., Bahou W. F., Docherty A. J., Cao J. Tissue inhibitor of metalloproteinase-2 (TIMP-2) binds to the catalytic domain of the cell surface receptor, membrane type 1-matrix metalloproteinase 1 (MT1-MMP). J Biol Chem. 1998;273(2):1216-1222.

    Not in our library10.1074/jbc.273.2.1216

39. 39.

    van Muijen G. N., Jansen K. F., Cornelissen I. M., Smeets D. F., Beck J. L., Ruiter D. J. Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice. Int J Cancer. 1991;48(1):85-91.

    Not in our library10.1002/ijc.2910480116

40. 40.

    van den Oord J. J., Paemen L., Opdenakker G., de Wolf-Peeters C. Expression of gelatinase B and the extracellular matrix metalloproteinase inducer EMMPRIN in benign and malignant pigment cell lesions of the skin. Am J Pathol. 1997;151(3):665-670.

    Not in our library

Cited by papers we hold 1

• Transmigration of Melanoma Cells through the Blood-Brain Barrier: Role of Endothelial Tight Junctions and Melanoma-Released Serine Proteases · 2011
  No notice

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.