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. 1999 Nov;81(6):934–941. doi: 10.1038/sj.bjc.6690790

A comparative evaluation of various invasion assays testing colon carcinoma cell lines

N J de Both 1, M Vermey 1, W N Dinjens 1, F T Bosman 2
PMCID: PMC2362956  PMID: 10576648

Abstract

Various colon carcinoma cell lines were tested in different invasion assays, i.e. invasion into Matrigel, into confluent fibroblast layers and into chicken heart tissue. Furthermore, invasive capacity and metastatic potential were determined in nude mice. The colon carcinoma cells used were the human cell lines Caco-2, SW-480, SW-620 and HT-29, and the murine lines Colon-26 and -38. None of the human colon carcinoma cells migrated through porous membranes coated with Matrigel; of the murine lines, only Colon-26 did. When incubated in a mixture of Matrigel and culture medium non-invading cells formed spheroid cultures, whereas invading cells showed a stellate outgrowth. Only the heterogeneously shaped (epithelioid and stellate) cells of SW-480 and SW-620 and the spindle-shaped cells of Colon-26 invaded clearly confluent skin and colon fibroblasts as well as chicken heart tissue. However, when transplanted into the caecum of nude and syngeneic mice, all the lines tested were invasive with the exception of Caco-2 cells. We conclude that the outcome of in vitro tests measuring the invasive capacity of neoplastic cells is largely dependent on the test system used. Invasive capacity in vitro is strongly correlated with cells having a spindle cell shape, vimentin expression and E-cadherin down regulation. In contrast, HT-29 and Colon-38 cells having an epithelioid phenotype were clearly invasive and metastatic in vivo, but not in vitro. © 1999 Cancer Research Campaign

Keywords: Matrigel, confluent fibroblast layers, chicken hearts, orthotopic transplantation

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Selected References

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  1. Albini A., Iwamoto Y., Kleinman H. K., Martin G. R., Aaronson S. A., Kozlowski J. M., McEwan R. N. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res. 1987 Jun 15;47(12):3239–3245. [PubMed] [Google Scholar]
  2. Boyer B., Vallés A. M., Thiery J. P. Model systems of carcinoma cell dispersion. Curr Top Microbiol Immunol. 1996;213(Pt 1):179–194. doi: 10.1007/978-3-642-61107-0_11. [DOI] [PubMed] [Google Scholar]
  3. Bresalier R. S., Raper S. E., Hujanen E. S., Kim Y. S. A new animal model for human colon cancer metastasis. Int J Cancer. 1987 May 15;39(5):625–630. doi: 10.1002/ijc.2910390514. [DOI] [PubMed] [Google Scholar]
  4. Chao C., Lotz M. M., Clarke A. C., Mercurio A. M. A function for the integrin alpha6beta4 in the invasive properties of colorectal carcinoma cells. Cancer Res. 1996 Oct 15;56(20):4811–4819. [PubMed] [Google Scholar]
  5. Corbett T. H., Griswold D. P., Jr, Roberts B. J., Peckham J. C., Schabel F. M., Jr Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays, with a note on carcinogen structure. Cancer Res. 1975 Sep;35(9):2434–2439. [PubMed] [Google Scholar]
  6. De Both N. J., Vermey M., Groen N., Dinjens W. N., Bosman F. T. Clonal growth of colorectal-carcinoma cell lines transplanted to nude mice. Int J Cancer. 1997 Sep 17;72(6):1137–1141. doi: 10.1002/(sici)1097-0215(19970917)72:6<1137::aid-ijc32>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  7. Ewing C. M., Ru N., Morton R. A., Robinson J. C., Wheelock M. J., Johnson K. R., Barrett J. C., Isaacs W. B. Chromosome 5 suppresses tumorigenicity of PC3 prostate cancer cells: correlation with re-expression of alpha-catenin and restoration of E-cadherin function. Cancer Res. 1995 Nov 1;55(21):4813–4817. [PubMed] [Google Scholar]
  8. Fabra A., Nakajima M., Bucana C. D., Fidler I. J. Modulation of the invasive phenotype of human colon carcinoma cells by organ specific fibroblasts of nude mice. Differentiation. 1992 Dec;52(1):101–110. doi: 10.1111/j.1432-0436.1992.tb00504.x. [DOI] [PubMed] [Google Scholar]
  9. Jeffers M., Rong S., Vande Woude G. F. Enhanced tumorigenicity and invasion-metastasis by hepatocyte growth factor/scatter factor-met signalling in human cells concomitant with induction of the urokinase proteolysis network. Mol Cell Biol. 1996 Mar;16(3):1115–1125. doi: 10.1128/mcb.16.3.1115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kath R., Jambrosic J. A., Holland L., Rodeck U., Herlyn M. Development of invasive and growth factor-independent cell variants from primary human melanomas. Cancer Res. 1991 Apr 15;51(8):2205–2211. [PubMed] [Google Scholar]
  11. Kinzler K. W., Vogelstein B. Lessons from hereditary colorectal cancer. Cell. 1996 Oct 18;87(2):159–170. doi: 10.1016/s0092-8674(00)81333-1. [DOI] [PubMed] [Google Scholar]
  12. Kunz-Schughart L. A., Kreutz M., Knuechel R. Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol. 1998 Feb;79(1):1–23. doi: 10.1046/j.1365-2613.1998.00051.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mareel M. M., Behrens J., Birchmeier W., De Bruyne G. K., Vleminckx K., Hoogewijs A., Fiers W. C., Van Roy F. M. Down-regulation of E-cadherin expression in Madin Darby canine kidney (MDCK) cells inside tumors of nude mice. Int J Cancer. 1991 Apr 1;47(6):922–928. doi: 10.1002/ijc.2910470623. [DOI] [PubMed] [Google Scholar]
  14. Mareel M., Kint J., Meyvisch C. Methods of study of the invasion of malignant C3H-mouse fibroblasts into embryonic chick heart in vitro. Virchows Arch B Cell Pathol Incl Mol Pathol. 1979 May 4;30(1):95–111. doi: 10.1007/BF02889094. [DOI] [PubMed] [Google Scholar]
  15. Meiners S., Brinkmann V., Naundorf H., Birchmeier W. Role of morphogenetic factors in metastasis of mammary carcinoma cells. Oncogene. 1998 Jan 8;16(1):9–20. doi: 10.1038/sj.onc.1201486. [DOI] [PubMed] [Google Scholar]
  16. Morikawa K., Walker S. M., Nakajima M., Pathak S., Jessup J. M., Fidler I. J. Influence of organ environment on the growth, selection, and metastasis of human colon carcinoma cells in nude mice. Cancer Res. 1988 Dec 1;48(23):6863–6871. [PubMed] [Google Scholar]
  17. Morton R. A., Ewing C. M., Nagafuchi A., Tsukita S., Isaacs W. B. Reduction of E-cadherin levels and deletion of the alpha-catenin gene in human prostate cancer cells. Cancer Res. 1993 Aug 1;53(15):3585–3590. [PubMed] [Google Scholar]
  18. Noël A. C., Callé A., Emonard H. P., Nusgens B. V., Simar L., Foidart J., Lapiere C. M., Foidart J. M. Invasion of reconstituted basement membrane matrix is not correlated to the malignant metastatic cell phenotype. Cancer Res. 1991 Jan 1;51(1):405–414. [PubMed] [Google Scholar]
  19. Polette M., Gilles C., de Bentzmann S., Gruenert D., Tournier J. M., Birembaut P. Association of fibroblastoid features with the invasive phenotype in human bronchial cancer cell lines. Clin Exp Metastasis. 1998 Feb;16(2):105–112. doi: 10.1023/a:1006572204497. [DOI] [PubMed] [Google Scholar]
  20. 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 Jul 7;370(6484):61–65. doi: 10.1038/370061a0. [DOI] [PubMed] [Google Scholar]
  21. Sieuwerts A. M., Klijn J. G., Foekens J. A. Assessment of the invasive potential of human gynecological tumor cell lines with the in vitro Boyden chamber assay: influences of the ability of cells to migrate through the filter membrane. Clin Exp Metastasis. 1997 Jan;15(1):53–62. doi: 10.1023/a:1018436407280. [DOI] [PubMed] [Google Scholar]
  22. Sommers C. L., Heckford S. E., Skerker J. M., Worland P., Torri J. A., Thompson E. W., Byers S. W., Gelmann E. P. Loss of epithelial markers and acquisition of vimentin expression in adriamycin- and vinblastine-resistant human breast cancer cell lines. Cancer Res. 1992 Oct 1;52(19):5190–5197. [PubMed] [Google Scholar]
  23. Sparks A. B., Morin P. J., Vogelstein B., Kinzler K. W. Mutational analysis of the APC/beta-catenin/Tcf pathway in colorectal cancer. Cancer Res. 1998 Mar 15;58(6):1130–1134. [PubMed] [Google Scholar]
  24. Sunitha I., Meighen D. L., Hartman D. P., Thompson E. W., Byers S. W., Avigan M. I. Hepatocyte growth factor stimulates invasion across reconstituted basement membranes by a new human small intestinal cell line. Clin Exp Metastasis. 1994 Mar;12(2):143–154. doi: 10.1007/BF01753981. [DOI] [PubMed] [Google Scholar]
  25. Thompson E. W., Paik S., Brünner N., Sommers C. L., Zugmaier G., Clarke R., Shima T. B., Torri J., Donahue S., Lippman M. E. Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J Cell Physiol. 1992 Mar;150(3):534–544. doi: 10.1002/jcp.1041500314. [DOI] [PubMed] [Google Scholar]
  26. Vermeulen S. J., Bruyneel E. A., Bracke M. E., De Bruyne G. K., Vennekens K. M., Vleminckx K. L., Berx G. J., van Roy F. M., Mareel M. M. Transition from the noninvasive to the invasive phenotype and loss of alpha-catenin in human colon cancer cells. Cancer Res. 1995 Oct 15;55(20):4722–4728. [PubMed] [Google Scholar]
  27. Vleminckx K., Vakaet L., Jr, Mareel M., Fiers W., van Roy F. Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role. Cell. 1991 Jul 12;66(1):107–119. doi: 10.1016/0092-8674(91)90143-m. [DOI] [PubMed] [Google Scholar]
  28. Webb G., Baker M. S., Nicholl J., Wang Y., Woodrow G., Kruithof E., Doe W. F. Chromosomal localization of the human urokinase plasminogen activator receptor and plasminogen activator inhibitor type-2 genes: implications in colorectal cancer. J Gastroenterol Hepatol. 1994 Jul-Aug;9(4):340–343. doi: 10.1111/j.1440-1746.1994.tb01252.x. [DOI] [PubMed] [Google Scholar]
  29. de Vries J. E., Dinjens W. N., De Bruyne G. K., Verspaget H. W., van der Linden E. P., de Bruïne A. P., Mareel M. M., Bosman F. T., ten Kate J. In vivo and in vitro invasion in relation to phenotypic characteristics of human colorectal carcinoma cells. Br J Cancer. 1995 Feb;71(2):271–277. doi: 10.1038/bjc.1995.55. [DOI] [PMC free article] [PubMed] [Google Scholar]

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