Skip to main content
SpringerLink
Log in

Transcriptomic signature of cell lines isolated from canine mammary adenocarcinoma metastases to lungs

  • Original Article
  • Published:
Journal of Applied Genetics Aims and scope Submit manuscript

Abstract

Metastasis is a final step in the progression of mammary gland cancer, usually leading to death. Potentially, a molecular signature of metastasis can be defined via comparison of primary tumors with their metastases. Currently, there is no data in the literature regarding the molecular portrait of metastases in dogs and only few reports regarding human cancer. This is the first report describing the transcriptomic signature of canine cancer metastatic cells. Two adenocarcinoma cell lines isolated from the canine mammary gland (CMT-W1 and CMT-W2) were compared with cell lines isolated from their lung metastases (CMT-W1M and CMT-W2M) with regards to the following cytometric parameters: cell cycle, ploidy, Bcl-2 expression, susceptibility to induced apoptosis, and transcriptomic profile. Cytometric analyses revealed significant differences in cell cycle and antiapoptotic potential between the examined cells. Using oligonucleotide microarrays, we found 104 up-regulated genes in the metastatic cell line CMT-W1M and 21 up-regulated genes in the primary CMT-W1 cell line. We also found 83 up-regulated genes in the CMT-W2M cell line and only 21 up-regulated genes in the CMT-W2 cell line. Among the up-regulated genes in both metastatic cell lines, we found 15 common genes. These differently expressed genes are involved mainly in signal transduction, cell structure and motility, nucleic acid metabolism, developmental processing, and apoptosis (GHSR, RASSF1, ARF1GAP, WDR74, SMOC2,SFRP4, DIAPH1, FSCN1, ALX4, SNX15, PLD2, WNT7B, POU6F2, NKG7, andPOLR2F). Seven ofthem are involved in a cellular pathway dependent on ghrelin via growth hormone secretagogue receptor (GHSR). Our results suggest that this pathway may be essential for mammary cancer cells to have a metastatic potential.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Agathanggelou A, Cooper WN, Latif F, 2005. Roleof the Ras-association domain family 1 tumor suppressor gene in human cancers. Cancer Res 63: 1888–1893.

    Google Scholar 

  • Brinkhof B, Spee B, Rothuizen J, Penning LC, 2006. Development and evaluation of canine reference genes for accurate quantification of gene expression. Anal Biochem 356: 36–43.

    Article  CAS  PubMed  Google Scholar 

  • Cameron MD, Schmidt EE, Kerkvliet N, Nadkarni KV, Morris VL, Groom AC, et al. 2000. Temporal progression of metastasis in lung: cell survival, dormancy, and locationdependenceofmetastatic inefficiency. Cancer Res 60: 2541–2546.

    CAS  PubMed  Google Scholar 

  • Darzynkiewicz Z, Robinson JP, Crissman HA, 1994. Flow cytometry, 2nd ed. Part A. Series: Methods Cell Biol: 41.

  • Dexbury MS, Waseem T, Ito H, Robinson MK, Zinner MJ, Ashley SW, et al. 2003. Grelin promotes pancreatic adenocarcinoma cellular proliferation and invasiveness. Biochem Biophys Res Commun 309: 464–468.

    Article  Google Scholar 

  • Dorn CR, Taylor DON, Frye FL, Hibbard HH, 1968. Survey of animal neoplasms in Alemeda and Contra Costa Counties, California I: methodology and description of cases. J Natl Cancer Inst 40: 295–305.

    CAS  PubMed  Google Scholar 

  • Etschmann B, Wilcken, B, Stoevesand K, von der Schulenburg A, Sterner-Kock A, 2006. Selection of reference genes for quantitative real-time PCR analysis in canine mammary tumors using the GeNorm algorithm. Vet Pathol 4: 934–942.

    Article  Google Scholar 

  • Fidler IJ, 1973. Selection of successive tumour lines for metastasis. Nat New Biol 242: 148–149.

    CAS  PubMed  Google Scholar 

  • Fidler IJ, Kripke ML, 2003. Genomic analysis of primary tumors does not address the prevalence of metastatic cells in the population. Nature Genet 34: 23.

    Article  CAS  PubMed  Google Scholar 

  • Fox MH, Armstrong LW, Withrow SJ, Powers BE, LeRue SM, Straw RC, el al. 1990. Comparison of DNA aneuploidy of primary and metastatic spontaneous canine osteosarcoma. Cancer Res 50: 6176–6178.

    CAS  PubMed  Google Scholar 

  • Gil-Puig C, Blanco M, Garcia-Caballero T, Segura C, Perez-Fernardez R, 2002. Pit-1/GHF-1 and GH expression in the MCF-7 human breast adenocarcinoma cell line. J Endocrinol 173: 161–167.

    Article  CAS  PubMed  Google Scholar 

  • Gruvberger-Saal SK, Eden P, Ringnér M, Baldetorp B, Chebil G, Borg A, et al. 2004. Predicting continuous values of prognostic markers in breast cancer from microarray gene expression profiles, Mol Cancer Ther 3: 161–168.

    CAS  PubMed  Google Scholar 

  • Hynes OR, 2003. Metastatic potential: generic predisposition of the primary tumor or rare, metastatic variants — or both? Cell 113: 821–823.

    Article  CAS  PubMed  Google Scholar 

  • Jeffrey PL, Murray RE, Yeh AH, McNamara JF, Duncan RP, Francis GD, et al. 2005. Expression and function of the ghrelin axis, including a novel preproghrelin isoform, in human breast cancer tissues and cell lines. Endocr Relat Cancer 12: 839–850.

    Article  Google Scholar 

  • Kakiuchi S, Daigo Y, Tsunoda T, Yano S, Sone S, Nakamura Y, 2003. Genome-wide analysis of organ-preferential metastasis of human small cell lung cancer in mice. Mol Cancer Res 1: 485–499.

    CAS  PubMed  Google Scholar 

  • Kang Y, Siegel PM, Shu W, Drobnjak M, Kakonen SM, Cordon-Cardo C, et al. 2003. A mutagenic program mediating breast cancer metastasis to bone. Cancer Cell 3: 537–549.

    Article  CAS  PubMed  Google Scholar 

  • Korczak B, Robson IB, Lamarche C, Bernstein A, Kerbel RS, 1988. Genetic tagging of tumor cells with retrovirus vectors: clonal analysis of tumor growth and metastasis in vitro. Mol Cell Biol 8: 3143–3149.

    CAS  PubMed  Google Scholar 

  • Król M, Pawłowski KM, Skierski J, Rao NAS, Hellmen E, Mol JA, et al. 2009. Transcriptomic profile of two canine cancer cell lines with different proliferative and anti-apoptotic potential. J Physiol Pharmacol 60: 95–106.

    PubMed  Google Scholar 

  • Lee AY, He B, You L, Dadfarmay S, Xu Z, Maziers J, et al. 2004. Expression of the secreted frizzled-related protein gene family is downregulated in human mesothelioma. Oncogene 23: 6672–6676.

    Article  CAS  PubMed  Google Scholar 

  • Li X, Zheng H, Hara T, Takahasi H, Masuda S, Wang Z, et al. 2008. Aberrant expression of cortactin and fascin are effective markers for pathogenesis, invasion, metastasis and prognosis of gastric carcinomas. Int J Oncol 33: 69–79.

    PubMed  Google Scholar 

  • Liu P, Lu J, Cardoso WV, Vaziri C, 2008. The SPARC-related factor SMOC-2 promotes growth factor-induced cyclin D1 expression and DNA synthesisviaintegrin-linkedkinase(ILK). Mol Biol Cell 19: 248–261.

    Article  CAS  PubMed  Google Scholar 

  • Lu P, Prost S, Caldwell H, Tugwood JD, Betton GR, Harrison DJ, 2007. Microarray analysis of gene expression of mouse hepatocytes of different ploidy. Mamm Genome 18: 617–626.

    Article  CAS  PubMed  Google Scholar 

  • MacEwen EG, 1990. Spontaneous tumors in dogs and cats: models for the study of cancer biology and treatment. Cancer Metastasis Rev 9: 125–136.

    Article  CAS  PubMed  Google Scholar 

  • Mi H, Lazareva-Ulitsky B, Loo R, Kejariwal A, Vandergriff J, Rabkin S, et al. 2005. The PANTHER database of protein families, subfamilies, functions and pathways. Nucleic Acids Res 33: 284–288.

    Article  Google Scholar 

  • Minn AJ, Gupta GP, Siegel PM, Bos PD, Shu W, Giri DD, et al. 2005. Genes that mediate breast cancer metastasis to lung. Nature 436: 518–524.

    Article  CAS  PubMed  Google Scholar 

  • Mol JA, van Garderen E, Selman PJ, Wolfswinkel J, Rijnberk A, Rutteman GE, 1995. Growth hormone mRNA inmammary glandtumorsindogs and cats. J Clin Invest 95: 2028–2034.

    Article  CAS  PubMed  Google Scholar 

  • Oshlack A, Emsile D, Corcoran LM, Smyth GK, 2007. Normalization of boutique two-color microarrays with a high proportion of differentially expressed probes. Genome Biol 8(1): R2.

    Article  PubMed  Google Scholar 

  • Pawłowski KM, Król M, Majewska A, Badowska-Kozakiewicz A, Mol JA, Malicka E, et al. 2009. Comparison of cellular and tissue transcriptomic profiles in canine mammary tumor. J Physiol Pharmacol 60: 85–94.

    PubMed  Google Scholar 

  • Ramaswamy S, Ross KN, Lander ES, Golub TR, 2003. A molecular signature of metastasis in primary solid tumors. Nat Genet 33: 49–54.

    Article  CAS  PubMed  Google Scholar 

  • Rao NAS, van Wolferen ME, Gracanin A, Bhatti SFM, Król M, Holstege FC, et al. 2009. Gene expression profiles of progestin-induced canine mammary hyperplasia and spontaneous mammary tumors. J Physiol Pharmacol 60: 73–84.

    PubMed  Google Scholar 

  • Rao NAS, van Wolferen ME, van der Ham R, van Leenen D, Groot Koerkamp MJA, Holstege FCP, et al. 2008. cDNA microarray profiles of canine mammary tumor cell lines reveal deregulated pathways pertaining to their phenotype. Anim Genet 39: 333–345.

    Article  CAS  PubMed  Google Scholar 

  • Tarin D, Price JE, Kettlewell MG, Souter RG, Vass AC, Crossley B, 1984. Clinicopathological observations on metastasis in man studied in patients treated with peritoneovenous shunts. Br Med J 288: 749–751.

    Article  CAS  Google Scholar 

  • Tusher VG, Tibshirani R, Chu G, 2001. Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A, 98: 5116–5121.

    Article  CAS  PubMed  Google Scholar 

  • Wang W, Wyckoff JB, Frohlich VC, Oleynikov Y, Huttelmaier S, Zavadil J, et al. 2002. Single cell behavior in metastatic primary mammary tumors correlated with gene expression patterns revealed by molecular profiling. Cancer Res 62: 6278–6288.

    CAS  PubMed  Google Scholar 

  • Weigelt B, Glas AM, Lodewyk F, Wessels A, Witteveen AT, Peterse JL, et al. 2003. Gene expression profiles of primary breast tumors maintained in distant metastases. Proc Natl Acad Sci U S A 100: 15901–15905.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences (SGGW), ul, Nowoursynowska 159, 02-776, Warsaw, Poland

    M. Król, K. M. Pawłowski, A. Majewska & T. Motyl

  2. Department and Clinic of Obstetrics, Ruminant Diseases and Animal Health Care, Wrocław University of Environmental and Life Sciences, Wroclaw, Poland

    J. Polańska

  3. Department of Internal Medicine, University of Giessen Lung Center, Justus Liebig University, Giessen, Germany

    P. Turowski & R. E. Morty

  4. Cell Biology Department, National Medicines Institute, Warsaw, Poland

    J. Skierski

  5. Department of Biochemistry, Pharmacology and Toxicology, Wrocław University of Environmental and Life Sciences, Wroclaw, Poland

    M. Ugorski

  6. Department Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wrocław, Wroclaw, Poland

    M. Ugorski

Authors
  1. M. Król
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Polańska
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. M. Pawłowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. Turowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Skierski
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. Majewska
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M. Ugorski
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R. E. Morty
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. T. Motyl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. Motyl.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Król, M., Polańska, J., Pawłowski, K.M. et al. Transcriptomic signature of cell lines isolated from canine mammary adenocarcinoma metastases to lungs. J Appl Genet 51, 37–50 (2010). https://doi.org/10.1007/BF03195709

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03195709

Keywords

Search

Navigation