Cytotechnology

, Volume 46, Issue 1, pp 9–18 | Cite as

Identification of three gene candidates for multicellular resistance in colon carcinoma

  • Nicholas E. Timmins
  • Tina L. Maguire
  • Sean M. Grimmond
  • Lars K. Nielsen
Article

Abstract

Solid tumours display elevated resistance to chemo- and radiotherapies compared to individual tumour derived cells. This so-called multicellular resistance (MCR) phenomenon can only be partly explained by reduced diffusion and altered cell cycle status; even fast growing cells on the surface of solid tumours display MCR. Multicellular spheroids (MCS) recapture this phenomenon ex vivo and here we compare gene expression in exponentially growing MCS with gene expression in monolayer culture. Using an 18,664 gene microarray, we identified 42 differentially expressed genes and three of these genes can be linked to potential mechanisms of MCR. A group of interferon response genes were also up-regulated in MCS, as were a number of genes that that are indicative of greater differentiation in three-dimensional cultures.

Keywords

3D culture Differential gene expression MCR MCS Multicellular resistance Multicellular spheroid Tumour 

Abbreviation

3D

Three-dimensional

5FU

5-Fluorouracil

MCR

Multicellular resistance

MCS

Multicellular spheroid

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References

  1. Arany, I., Chen, S.H., Megyesi, J.K., Adler-Storthz, K., Chen, Z., Rajaraman, S., Ember, I.A., Tyring, S.K., Brysk, M.M. 2003Differentiation-dependent expression of signal transducers and activators of transcription (STATs) might modify responses to growth factors in the cancers of the head and neckCancer Lett.1998389CrossRefPubMedGoogle Scholar
  2. Baldwin, A.S. 2001Control of oncogenesis and cancer therapy resistance by the transcription factor NF-kappa BJ. Clin. Invest.107241246PubMedGoogle Scholar
  3. Courilleau, D., Chastre, E., Sabbah, M., Redeuilh, G., Atfi, A., Mester, J. 2000B-ind1, a novel mediator of Rac1 signaling cloned from sodium butyrate-treated fibroblastsJ. Biol. Chem.2751734417348CrossRefPubMedGoogle Scholar
  4. Debatin, K.M. 2004Apoptosis pathways in cancer and cancer therapyCancer Immunol. Immunother53153159CrossRefPubMedGoogle Scholar
  5. Desoize, B., Jardillier, J. 2000Multicellular resistance: a paradigm for clinical resistance?Crit. Rev. Oncol. Hematol.36193207PubMedGoogle Scholar
  6. Dubessy, C., Merlin, J.M., Marchal, C., Guillemin, F. 2000Spheroids in radiobiology and photodynamic therapyCrit. Rev. Oncol. Hematol.36179192PubMedGoogle Scholar
  7. Flatmark, K., Pedersen, K., Nesland, J., Rasmussen, H., Aamodt, G., Mikalsen, S., Bjornland, K., Fodstad, O., Maelandsmo, G. 2003Nuclear localization of the metastasis-related protein s100a4 correlates with tumour stage in colorectal cancerJ. Pathol.200589595CrossRefPubMedGoogle Scholar
  8. Fracasso, G., Colombatti, M. 2000Effect of therapeutic macromolecules in spheroidsCrit. Rev. Oncol. Hematol.36159178PubMedGoogle Scholar
  9. Hamilton, G. 1998Multicellular spheroids as an in vitro tumor modelCancer Lett.1312934CrossRefPubMedGoogle Scholar
  10. Huang, J., Teng, L., Liu, T., Li, L., Chen, D., Li, F., Xu, L.G., Zhai, Z., Shu, H.B. 2003Identification of a novel serine/threonine kinase that inhibits TNF-induced NF-kappaB activation and p53-induced transcriptionBiochem. Biophys. Res. Commun.309774778CrossRefPubMedGoogle Scholar
  11. Hwang, B.J., Ford, J.M., Hanawalt, P.C., Chu, G. 1999Expression of the p48 xeroderma pigmentosum gene is p53-dependent and is involved in global genomic repairProc. Natl. Acad. Sci. USA96424428CrossRefPubMedGoogle Scholar
  12. Kelm, J.M., Timmins, N.E., Brown, C.J., Fussenegger, M., Nielsen, L.K. 2003Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell typesBiotechnol. Bioeng.83173180CrossRefPubMedGoogle Scholar
  13. Knowles, H.J., Phillips, R.M. 2001Identification of differentially expressed genes in experimental models of the tumor microenvironment using differential displayAnticancer Res.2123052311PubMedGoogle Scholar
  14. Kunz-Schughart, L., Mueller-Klieser, W. 2000

    Three-dimensional culture

    Masters, J. eds. Animal Cell Culture: A Practical Approach. 3Oxford University PressNew York123184
    Google Scholar
  15. Kunz-Schughart, L.A., Kreutz, M., Knuechel, R. 1998Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biologyInt. J. Exp. Pathol.79123CrossRefPubMedGoogle Scholar
  16. Lazareff, J.A., Suwinski, R., De Rosa, R., Olmstead, C.E. 1999Tumor volume and growth kinetics in hypothalamic-chiasmatic pediatric low grade gliomasPediatr. Neurosurg.30312319CrossRefPubMedGoogle Scholar
  17. Luker, K., Pica, C., Schreiber, R., Piwnica-Worms, D. 2001Overexpression of irf9 confers resistance to antimicrotubule agents in breast cancer cellsCancer Res.6165406547PubMedGoogle Scholar
  18. Marusic, M., Bajzer, Z., Vuk-Pavlovic, , Freyer, J. 1994Tumour growth in vivoas multicellular spheroids compared by mathematical modelsBull. Math. Biol.56617631CrossRefPubMedGoogle Scholar
  19. Mayo, M.W., Baldwin, A.S. 2000The transcription factor NF-kappaB: control of oncogenesis and cancer therapy resistanceBiochim. Biophys. Acta1470M5562PubMedGoogle Scholar
  20. Moriyama-Kita, M., Endo, Y., Yonemura, Y., Heizmann, C., Schafer, B., Sasaki, T., Yamamoto, E. 2004Correlation of s100a4 expression with invasion and metastasis in oral squamous cell carcinomaOral Oncol.40496500CrossRefPubMedGoogle Scholar
  21. Mueller-Klieser, W. 1997Three-dimensional cell cultures: from molecular mechanisms to clinical applicationsAm. J. Physiol.273C11091123PubMedGoogle Scholar
  22. Nederman, T., Twentyman, P. 1984Spheroids for studies of drug effectsRecent Results Cancer Res.9584102PubMedGoogle Scholar
  23. Olive, P.L., Durand, R.E. 1994Drug and radiation resistance in spheroids: cell contact and kineticsCancer Metastasis Rev.13121138CrossRefPubMedGoogle Scholar
  24. Olive, P.L., Banath, J.P., Evans, H.H. 1993Cell killing and DNA damage by etoposide in Chinese hamster V79 monolayers and spheroids: influence of growth kinetics, growth environment and DNA packagingBr. J. Cancer67522530PubMedGoogle Scholar
  25. Oloumi, A., MacPhail, S.H., Johnston, P.J., Banath, J.P., Olive, P.L. 2000Changes in subcellular distribution of topoisomerase IIalpha correlate with etoposide resistance in multicell spheroids and xenograft tumorsCancer Res.6057475753PubMedGoogle Scholar
  26. Oloumi, A., Lam, W., Banath, J.P., Olive, P.L. 2002Identification of genes differentially expressed in v79 cells grown as multicell spheroidsInt. J. Radiation Biol.78483492CrossRefGoogle Scholar
  27. Perou, C.M., Jeffrey, S.S., Rijn, M., Rees, C.A., Eisen, M.B., Ross, D.T., Pergamenschikov, A., Williams, C.F., Zhu, S.X., Lee, J.C., Lashkari, D., Shalon, D., Brown, P.O., Botstein, D. 1999Distinctive gene expression patterns in human mammary epithelial cells and breast cancersProc. Natl. Acad. Sci. USA9692129217CrossRefPubMedGoogle Scholar
  28. Poland, J., Sinha, P., Siegert, A., Schnolzer, M., Korf, U., Hauptmann, S. 2002Comparison of protein expression profiles between monolayer and spheroid cell culture of ht-29 cells revealed fragmentation of ck18 in three-dimensional cell cultureElectrophoresis2311741184CrossRefPubMedGoogle Scholar
  29. Rainaldi, G., Calcabrini, A., Arancia, G., Santini, M.T. 1999Differential expression of adhesion molecules (CD44, ICAM-1 and LFA-3) in cancer cells grown in monolayer or as multicellular spheroidsAnticancer Res.1917691778PubMedGoogle Scholar
  30. Ramakers, C., Ruijter, J.M., Deprez, R.H., Moorman, A.F. 2003Assumptionfree analysis of quantitative real-time polymerase chain reaction (PCR) dataNeurosci. Lett.3396266CrossRefPubMedGoogle Scholar
  31. Rosty, C., Ueki, T., Argani, P., Jansen, M., Yeo, C., Cameron, J., Hruban, R., Goggins, M. 2002Overexpression of S100A4 in pancreatic ductal adenocarcinomas is associated with poor differentiation and dna hypomethylationAm. J. Pathol.1604550PubMedGoogle Scholar
  32. Shiras, A., Bhosale, A., Patekar, A., Shepal, V., Shastry, P. 2002Differential expression of CD44(S) and variant isoforms v3, v10 in three-dimensional cultures of mouse melanoma cell linesClin. Exp. Metastasis19445455CrossRefPubMedGoogle Scholar
  33. Sutherland, R. 1988Cell and environment interactions in tumor microregions: the multicell spheroids modelScience240177184PubMedGoogle Scholar
  34. Wartenberg, M., Frey, C., Diedershagen, H., Ritgen, J., Hescheler, J., Sauer, H. 1998Development of an intrinsic P-glycoprotein-mediated doxorubicin resistance in quiescent cell layers of largemulticellular prostate tumor spheroidsInt. J. Cancer75855863CrossRefPubMedGoogle Scholar
  35. Weaver, V.M., Lelievre, S., Lakins, J.N., Chrenek, M.A., Jones, J.C., Giancotti, F., Werb, Z., Bissell, M.J. 2002Beta4 integrin-dependent formation of polarized three-dimensional architecture confers resistance to apoptosis in normal and malignant mammary epitheliumCancer Cell2205216CrossRefPubMedGoogle Scholar
  36. Wu, M., Xu, L.G., Zhai, Z., Shu, H.B. 2003SINK is a p65-interacting negative regulator of NF-kappaB-dependent transcriptionJ. Biol. Chem.2782707227079CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Nicholas E. Timmins
    • 1
  • Tina L. Maguire
    • 2
  • Sean M. Grimmond
    • 2
  • Lars K. Nielsen
    • 1
  1. 1.Laboratory for Biological Engineering, Department of Chemical EngineeringUniversity of QueenslandBrisbaneAustralia
  2. 2.The Institute for Molecular BiosciencesThe University of QueenslandBrisbaneAustralia

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