Clonal B16-F10 cell lines with increased expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) have been generated by transfection with a TIMP-1-containing expression vector. The parental B16-F10 and control 1–2 cells, and two TIMP-1 up-regulated clones (2–10, 6–5), were studied for their growth characteristics in tissue culture and their experimental metastatic ability in the chick embryo. Both of the TIMP-1 up-regulated clones showed slower in vitro growth and had lower saturation densities. Both clones were also less metastatic following intravenous injection into chick embryos, and formed significantly fewer metastatic tumors in the chorioallantoic membrane and in the liver than did parental B16-F10 and control cells. Furthermore, the size of tumors formed by TIMP-1 up-regulated cells was significantly reduced in comparison to the tumors produced by B16-F10 or control cells. Our results show that malignant cell lines genetically modified to express increased levels of TIMP-1 exhibit a suppressed experimental metastatic ability in vivo. We propose that TIMP-1 suppresses metastatic ability by decreasing both invasive and growth abilities.
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References
Liotta LA, Steeg PS and Stetler-Stevenson WG, 1991, Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell, 64, 327–336.
Khokha R and Denhardt DT, 1989, Matrix metalloproteinases and tissue inhibitor of metalloproteinases: a review of their role in tumorigenesis and tissue invasion. Invasion and Metastasis, 9, 391–405.
Liotta LA, 1990, Introductory overview. Cancer and Metastasis Reviews, 9, 285–287.
Halaka AN, Bunning RA, Bird CC, Gibson M and Reynolds JJ, 1983, Production of collagenase and inhibitor (TIMP) by intracranial tumors and dura in vitro. Journal of Neurosurgery, 49, 461–466.
Hicks NJ, Ward RV and Reynolds JJ, 1984, A fibrosarcoma model derived from mouse embryo cells: growth properties and secretion of collagenase and metalloproteinase inhibitor (TIMP) by tumor cell lines. International Journal of Cancer, 33, 835–844.
Mignatti P, Robbins E and Rifkin DB, 1986, Tumor invasion through the human amniotic membrane: requirement for a protein cascade. Cell, 47, 487–498.
Gavriolic J, Hembry RM, Reynolds JJ and Murphy G, 1987, Tissue inhibitor of metalloproteinases (TIMP) regulated extracellular type I collagen degradation by chondrocytes and endothelial cells. Journal of Cell Science, 87, 357–362.
Lala PK and Graham CH, 1990, Mechanisms of trophoblast invasiveness and their control: the role of proteases and protease inhibitors. Cancer and Metastasis Reviews, 9, 369–379.
Shultz RM, Silberman S, Persky B, Bajkowski AS and Carmichael DF, 1988, Inhibition by human recombinant tissue inhibitor of metalloproteinases of human amnion invasion and lung colonization by murine B16-F10 melanoma cells. Cancer Research, 48, 5539–5545.
Khokha R, Waterhouse P, Yagel S, Lala PK, Overall CM, Norton G and Denhardt DT, 1989, Antisense RNA-induced reduction in murine TIMP levels confers oncogenicity on Swiss 3T3 cells. Science, 243, 947–950.
Khokha R, Zimmer MJ, Graham CH, Lala PK and Waterhouse P, 1992, Suppression of invasion by inducible expression of tissue inhibitor of metallo proteinase-1 (TIMP-1) in B16-F10 melanoma cells Journal of the National Cancer Institute, 84, 1017–1022.
Chambers AF, Shafir R and Ling V, 1982, A model system for studying metastasis using the embryonic chick. Cancer Research, 42, 4018–4025.
Chambers AF, Wilson SM, Tuck AB, Denhardt GH and Cairncross JG, 1990, Comparison of metastatic properties of a variety of mouse, rat, and human cells in assays in nude mice and chick embryos. In Vivo, 4, 215–220.
Fidler IJ, 1975, Biological behavior of malignant cells correlated to their survival in vivo. Cancer Research, 35, 218–224.
Khokha R and Denhardt DT, 1987, On the use of anti-sense RNA: down-regulation of mRNA encoding a metalloproteinase inhibitor. Anticancer Research, 7, 653–660.
Chambers AF, Denhardt GH and Wilson SM, 1990, Ras-transformed NIH 3T3 cell lines, selected for metastatic ability in chick embryos, have increased proportions of p21-expressing cells and are metastatic in nude mice. Invasion and Metastasis, 10, 225–240.
Albini A, Melchiori A, Santi L, Liotta LA, Brown PD and Stetler-Stevenson WG, 1991, Tumor cell invasion inhibited by TIMP-2. Journal of the National Cancer Institute, 83, 775–779.
DeClerck YA, Yean TD, Chan D, Shimada H and Langley KE, 1991, Inhibition of tumor invasion of smooth muscle cell layers by metalloproteinase inhibitor (rMl/rTIMP-2). Cancer Research, 51, 2151–2157.
Denhardt DT, Khokha R, Yagel S, Overall CM and Parhar RS. Oncogenic consequences of downmodulating TIMP expression in 3T3 cells with anti sense RNA. In: Birkedal-Hansen H, Werb Z, Welgus HG and Van Wart HE, eds. Matrix Supplement No. 1, pp. 281–285 New York: Gustav Fischer Verlag, 1992.
Moses MA, Sudhalter J and Langer R, 1990, Identification of an inhibitor of neovascularization from cartilage. Science, 248, 1408–1410.
Takigawa M, Yukimitsu N, Suzuki F, Kishi J-I, Yamashita K and Hayakawa T, 1990, Induction of angiogenesis in chick yolk-sac membrane by poly amines and its inhibition by tissue inhibitors of metalloproteinases (TIMP and TIMP-2). Biochemical and Biophysical Research Communications, 171, 1264–1271.
Bertaux B, Hornebeck W, Eisen AZ and Duberret L, 1991, Growth stimulation of human keratinocytes by tissue inhibitor of metalloproteinases. Journal of Investigative Dermatology, 97, 679–685.
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Khokha, R., Zimmer, M.J., Wilson, S.M. et al. Up-regulation of TIMP-1 expression in B16-F10 melanoma cells suppresses their metastatic ability in chick embryo. Clin Exp Metast 10, 365–370 (1992). https://doi.org/10.1007/BF00133464
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DOI: https://doi.org/10.1007/BF00133464