Overview of Metastasis Assays

Cloud P. Paweletz1, Lu Charboneau1, Lance A. Liotta1

1 National Cancer Institute, NIH, Bethesda, Maryland
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 19.1
DOI:  10.1002/0471143030.cb1901s12
Online Posting Date:  November, 2001
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During tumor progression, cells acquire genetic and proteomic changes as they transform from normal to hyperplastic, through dysplasia, to carcinoma in situ, and finally to invasive and metastatic. The time course of progression may extend as far back as 10 years prior to diagnosis. Discerning the mechanism whereby tumor cells execute metastatic dissemination may provide the foundation necessary for successful treatment of the disease. For example, direct genetic evidence has linked in situ breast cancer to invasive carcinoma of the breast supporting the generally accepted assumption that carcinoma in situ of the breast is a clonal expansion of hyperproliferating cells. This in turn may provide a more comprehensive and/or functionally directed target strategy for intervention and prevention of breast cancer. This overview provides a picture of the processes related to metastasis and the experimental approaches used to study these processes.

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Table of Contents

  • Literature Cited
  • Figures
  • Tables
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Literature Cited

Literature Cited
  Anzano, M.A., Roberts, A.B., Smith, J.M., Sporn, M.B., and De Larco, J.E. 1983. Sarcoma growth factor from conditioned medium of virally transformed cells is composed of both type alpha and type beta transforming growth factors. Proc. Natl. Acad. Sci. U.S.A. 80:6264‐6268.
  Astrow, A.B. 1994. Commentary: Rethinking cancer. Lancet 343:494‐495.
  Axelrod, J.H., Reich, R., and Miskin, R. 1989. Expression of human recombinant plasminogen activators enhance invasion and experimental metastasis of H‐ras‐transformed NIH 3T3 cells. Mol. Cell. Biol. 9:2133‐2141.
  Behrens, J., Mareel, M.M., van Roy, F.M., and Birchmeier, W. 1989. Dissecting tumor cell invasion: Epithelial cells acquire invasive properties after the loss of uvomorulin‐mediated cell‐cell adhesion. J. Cell Biol. 108:2435‐2447.
  Chambers, A.F. and Matrisian, L.M. 1997. Changing views of the role of matrix metalloproteinases in metastasis. J. Natl. Cancer Inst. 89:1260‐1270.
  Chammas, R. and Brentani, R. 1991. Integrins and metastases: An overview. Tumour Biol. 12:309‐320.
  Condeelis, J. 1993. Life at the leading edge: The formation of cell protrusion. Annu. Rev. Physiol. 9:411‐444.
  Damsky, C.H. and Werb, Z. 1992. Signal transaction by integrin receptors for extracellular matrix: Cooperation and processing of extracellular information. Curr. Opin. Cell Biol. 4:772‐781.
  DeClerck, Y.A., Yean, T.D., Chan, D., Shimada, H., and Langley, K.E. 1991. Inhibition of tumor cell invasion of smooth muscle cell layers by recombinant human metalloproteinase inhibitor. Cancer Res. 51:2151‐2157.
  Di Renzo, M.F., Poulsom, R., Olivero, M., Comoglio, P.M., and Lemoine, N.R. 1995. Expression of the Met/hepatocyte growth factor receptor in human pancreatic cancer. Cancer Res. 55:1129‐1138.
  Dvorak, H.F., Brown, L.F., Detmar, M., and Dvorak, A.M. 1995. Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am. J. Path. 146:1029‐1039.
  Emmert‐Buck, M.R., Gillespie, J.W., Paweletz, C.P., Ornstein, D.K., Basrur, V., Appella, E., Wang, Q.‐H., Huang, J., Hu, N., Taylor, P., and Petricoin, E.F. 2000. An approach to proteomic analysis of human tumors. Mol. Carcinog. 27:158‐165.
  Emonard, H. and Grimaud, J.A. 1990. Matrix metalloproteinases. A review. Cell. Mol. Biol. 36:131‐153.
  Esheicher, A., Wohlwend, A., Belin, D., and Vassalli, J.D. 1989. Characterization of the cellular binding site for the urokinase type plasminogen activator. J. Biol. Chem. 264:1180‐1189.
  Farina, K.L., Wyckoff, J.B., Rivera, J., Lee, H., Segall, J.E., Condeelis, J.S., and Jones, J.G. 1998. Cell motility of tumor cells visualized in living intact primary tumors using green fluorescent protein. Cancer Res. 58:2528‐2532.
  Fidler, I.J. 1991. New developments in in vivo models of neoplasia. Cancer Metastasis Rev. 10:191‐192.
  Fidler, I.J. and Ellis, L.M. 1994. The implication of angiogenesis for the biology and therapy of cancer metastasis. Cell 79:185‐188.
  Fidler, I.J. and Hart, I.R. 1982. Biologic diversity in metastatic neoplasms–origins and implications. Science 217:998‐1003.
  Folkman, J. 1995. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat. Med. 1:27‐31.
  Folkman, J. and Klagsbrun, M. 1987. Angiogenic factors. Science 235:442‐447.
  Frixen, U.H., Behrens, J., Sachs, M., Eberle, G., Voss, B., Warda, A., Lochner, D., and Birchmeier, W. 1991. E‐cadherin mediated cell‐cell adhesion prevents invasiveness of human carcinoma cells. J. Cell Biol. 113:173‐185.
  Gallager, H.S. and Martin, J.E. 1969. The study of mammary carcinoma by mammography and whole organ sectioning, early observation. Cancer 23:855‐873.
  Granovsky, M., Fata, J., Pawling, J., Muller, W.J., Khokha, R., and Dennis, J.W. 2000. Suppression of tumor growth and metastasis in Mgat5‐deficient mice. Nat. Med. 6:306‐312.
  Guan, R.J., Ford, H.L., Fu, Y., Li, Y., Shaw, L.M., and Pardee, A.B. 2000. Drg‐1 as a differentiation‐related, putative metastatic suppressor gene in human colon cancer. Cancer Res. 60:749‐755.
  Guirguis, R., Margulies, I., Taraboletti, G., Schiffmann, E., and Liotta, L.A. 1987. Cytokine induced pseudopodial protrusion is coupled to tumor metastasis. Nature 329:261‐263.
  Hullet, M.D., Freeman, C., Hamdorf, B.J., Baker, R.T., Harris, M.J., and Parish, C.R. 1999. Cloning of mammalian heparinase, an important enzyme in tumor invasion and metastasis. Nat. Med. 5:735‐736.
  Hynes, R.O. 1992. Integrins: Versatility, modulation, and signalling in cell adhesion. Cell 69:11‐25.
  Juliano, R.L. and Haskill, S. 1993. Signal transduction from the extracellular matrix. J. Cell Biol. 120:577‐585.
  Kerbel, R.S., Cornil, I., and Theodorescu, D. 1991. Importance of orthotopic procedures in assessing the effects of transfected genes on human tumor growth and metastasis. Cancer Metastasis Rev. 10:201‐215.
  Koblinski, E.J., Ahram, M., and Sloane, B.F. 2000. Unraveling the role of proteases in cancer. Clin. Chim. Acta 291:113‐135.
  Lawson, R.S., Latter, G., Miller, S.D., Goldstein, D., Naps, M., Burbeck, S., Teng, N.N.H., and Zuckerkandl, E. 1991. Quantitative protein changes in metastatic versus primary epithelial ovarian carcinoma. Gynecol. Oncol. 41:22‐27.
  Levy, A., Cioce, V., Sobel, M.E., Garbisa, S., Grigioni, W.F., Liotta, L.A., and Stetler‐Stevenson, W.G. 1991. Increased expression of the 72 kDa type IV collagenase in human colonic adenocarcinoma. Cancer Res. 51:439‐444.
   Liotta, L.A. and Stetler‐Stevenson, W.G. 1993. Principles of molecular cell biology of cancer: Cancer metastasis. In Cancer: Principles and Practices of Oncology, Fourth edition (V.T. deVita, Jr., S. Hellman, and S.A. Rosenberg, eds.) pp.134‐149. Lippincott, Philadelphia.
  Liotta, L.A., Kleinerman, J., and Saidel, G. 1974. Quantitative relationships of intravascular tumor cells: Tumor vessels and pulmonary metastases following tumor implatation. Cancer Res. 34:997‐1004.
  Liotta, L.A., Tryggvason, K., Garbisa, S., Hart, I., Foltz, C.M., and Shafie, S. 1980. Metastatic potential correlates with enzymatic degradation of basement membranes. Nature 284:67‐68.
  Liotta, L.A., Steeg, P.S., and Stetler‐Stevenson, W.G. 1991. Cancer metastasis and angiogenesis; an imbalance of positive and negative regulation. Cell 68:327‐336.
  Mareel, M.M., Behrens, J., Birchmeier, W., De Bruyne, G.K., Vleminckx, K., Hoogewijs, A., Fiers, W.C., and van Roy, F.M. 1991. Down regulation of E‐cadherin expression in Madin Darby canine kidney (MDCK) cell tumors of nude mice. Int. J. Cancer 47:922‐928.
  Mignatti, P., Tsuboi, R., Robbins, E., and Rifkin, D. 1989. In vitro angiogenesis on the human amniotic membrane: Requirement for basic fibroblast growth factor‐induced proteinases. J. Cell Biol. 108:671‐682.
  Monteagudo, C., Merino, M., San Juan, J., Liotta, L.A., and Stetler‐Stevenson, W.G. 1990. Immunohistologic distribution of type IV collagenases in normal, benign, and malignant breast tissue. Am. J. Path. 136:585‐592.
  Nabeshima, K., Inoue, T., Shimao, Y., Kataoka, H., and Koono, M. 1999. Cohort migration of carcinoma cells: Differentiated colorectal cells move as coherent cell clusters or sheets. Histol. Histopathol. 14:1183‐1197.
  Nakajima, M., Welch, D., Belloni, P.N., and Nicolson, G.L. 1987. Degradation of basement membrane type IV collagen and lung subendothelial matrix by rat mammary adenocarcinoma cell clones of differing metastatic potentials. Cancer Res. 47:4869‐4876.
  Nakajima, M., Irimura, T., and Nicolson, G.L. 1988. Heparanase and tumor metastasis. J. Cell. Biochem. 36:157‐167.
  Nam, S.W., Clair, T., Campo, C.K., Lee, H.Y., Liotta, L.A., and Stracke, M.L. 2000. Autotaxin (ATX), a potent tumor motogen, augments invasive and metastatic potential of ras‐transformed cells. Oncogene 19:241‐247.
  Natali, P.G., Nictora, M.R., Cavaliere, R., Giannarelli, D., and Bigotti, A. 1991. Tumor progression in human malignant melanoma is associated with changes in α6/β1 laminin receptor. Int. J. Cancer 49:168‐172.
  Nicolson, G.L. 1988. Organ specificity of tumor metastasis: Role of preferential adhesion, invasion, and growth malignant cells at specific secondary sites. Cancer Metastasis Rev. 7:143‐188.
  Nicolson, G.L. 1991. Gene expression, cellular diversification and tumor progression to the metastatic phenotype. BioEssays 13:337‐342.
  Ossowski, L. and Reich, E. 1983. Antibodies to plasminogen activator inhibit human tumor metastasis. Cell 35:611‐619.
  Ostrowski, L.E., Rinch, J., King, P., and Matrisian, L. 1988. Expression pattern of a gene for a secreted metalloproteinase during late stages of tumor progression. Mol. Carcinog. 1:13‐19.
  Parish, C.R., Coombe, D.R., Jakobsen, K.B., and Underwood, P.A. 1987. Evidence that sulphated polysaccharides inhibit tumor metastasis by blocking tumor cell‐derived heparanase. Int. J. Cancer 40:511‐518.
  Reich, R., Thompson, E., Iwamoto, Y., Martin, G.R., Deason, J.R., Fuller, G.C., and Miskin, R. 1988. Effects of inhibitors of plasminogen activator, serine proteases, and collagenase IV on the invasion of basement membranes by metastatic cells. Cancer Res. 48:3307‐3312.
  Spratt, J.S., Greenberg, R.A., and Heuser, L.S. 1986. Geometry, growth rates, and duration of cancer and carcinoma in situ of the breast before detection by screening. Cancer Res. 46:970‐974.
  Stossel, T.P. 1993. On the crawling of animal cells. Science 260:1086‐1094.
  Strausberg, R.L., Dahl, C.A., and Klausner, R.D. 1997. New opportunities for uncovering the molecular basis of cancer. Nat. Genet. 15:415‐416.
  Takeichi, M. 1991. Cadherin cell adhesion receptors as a morphogenetic regulator. Science 251:1451‐1455.
  Ura, H., Bonfil, R.D., Reich, R., Reddel, R., Pfeifer, A., Harris, C.C., and Klein‐Szanto, A.J. 1989. Expression of type IV collagenase and procollagen genes and its correlation with tumorigenic, invasive, and metastatic abilities of oncogene transformed human bronchial cells. Cancer Res. 49:4615‐4621.
  Vleminckx, K., Vakaet, L. Jr., Fiers, W., and Van Roy, F.M. 1991. Genetic manipulation of E‐cadherin by epithelial tumor cells reveals an invasion suppressor role. Cell 66:107‐119.
  Vlodavsky, I., Mohsen, M., Lider, O., Svahn, C.M., Ekre, H.P., Vigoda, M., Ishai‐Michaeli, R., and Peretz, T. 1995. Inhibition of tumor metastasis by heparanase inhibiting species of heparin. Invasion Metastasis 14:290‐302.
  Vlodavsky, I., Friedmann, Y., Elkin, M., Aingorn, H., Atzmon, R., Ishai‐Michaeli, R., Bitan, M., Pappo, O., Peretz, T., Miichal, I., Spector, L., and Pecker, I. 1999. Mammalian heparanase: Gene cloning, expression and function in tumor progression and metastasis. Nat. Med. 5:793‐802.
  You, J., Aznavoorian, S., Liotta, L.A., and Dong, C. 1996. Responses of tumor cell pseudopod protrusion to changes in medium osmolality. J. Cell Physiol. 167:156‐163.
  Zhuang, Z., Merino, M., Chuaqui, R., Liotta, L.A., and Emmert‐Buck, M. 1995. Identical allelic loss on chromosome 11q13 in microdissected in situ and invasive human breast cancer. Cancer Res. 55:467‐471.
  Zuber, J., Tchernitsa, O.I., Hinzmann, B., Schmitz, A.C., Grips, M., Hellriegel, M., Sers, C., Rosenthal, A., and Schafer, R. 2000. A genome‐wide survey of RAS transformation targets. Nature Genet. 24:144‐152.
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