Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Open Life Sciences

formerly Central European Journal of Biology

Editor-in-Chief: Ratajczak, Mariusz

1 Issue per year

IMPACT FACTOR 2017: 0.764
5-year IMPACT FACTOR: 0.787

CiteScore 2017: 0.88

SCImago Journal Rank (SJR) 2017: 0.271
Source Normalized Impact per Paper (SNIP) 2017: 0.545

Open Access
See all formats and pricing
More options …
Volume 2, Issue 1


Volume 10 (2015)

Changes in cytokine production and morphology of murine lymphoma NK/Ly cells in course of tumor development

Rostyslav Panchuk / Natalia Boiko / Maxim Lootsik / Rostyslav Stoika
Published Online: 2007-03-01 | DOI: https://doi.org/10.2478/s11535-007-0011-4


The main goal of this study was to evaluate if specific cytokine expression in the NK/Ly lymphoma cells might be involved in development of intoxication in the tumor-bearing animals. RT-PCR analysis was used to study an expression of mRNA coding for IL-1α, IL-6, TNF-α, TNF-β and VEGF. ELISA was used to evaluate IL-6 and IFN-γ concentration in the ascitic fluid. Cytomorphological investigation of tumor cells was done after standard Romanovsky-Giemsa staining, and chromatin staining was performed with hematoxyline and neutral red. Lactate dehydrogenase and acid phosphatase release from tumor cells was estimated. It was revealed that the level of mRNA coding for VEGF and IL-6 was significant in the lymphoma cells. The level of VEGF mRNA was initially high and did not change during tumor progression, while the level of expression of IL6 mRNA was low at the initial stages of tumor growth and markedly increased (up to 5-fold) at the terminal stages. The obtained data on IL-6 mRNA expression were confirmed by ELISA, which showed more than 6-fold increase (from 90 to 570 pg/ml) in the IL-6 concentration in the ascitic fluid at late stages of NK/Ly tumor development. On the contrary to IL-6, concentration of IFN-γ in the ascitic fluid was very high at early stages of tumor development (1,000 pg/ml) and it markedly decreased (up to 30-fold, 30 pg/ml) at the terminal stages of tumor development. The high levels of IL-6 mRNA in tumor cells and IL-6 content in extracellular medium correlated with cell deterioration, as revealed by cytomorphologic study and the release of intracellular enzymes into extracellular medium. We suggest that an enhanced production and release of IL-6 by lymphoma cells can cause intoxication and exhaustion of the organism observed at terminal stages of tumor growth.

Keywords: Murine lymphoma NK/Ly; cytokines; tumor ageing

  • [1] L. Nemeth and B. Kellner: “A new mouse ascites tumor to be used as screening tool”, Neoplasma, Vol. 8, (1961), pp. 337–343. Google Scholar

  • [2] L. Kopper, A. Jeney, J. Takács, I. Benedeczky, E. Dzurillay and K. Lapis: “Studies of the Growth of an Ascitic Tumour. I. Cellular and Subcellular Changes During the Tumour Growth”, Eur. J. Cancer, Vol. 14, (1978), pp. 59–73. CrossrefGoogle Scholar

  • [3] L. Kopper, A. Jeney, K. Lapis and M. Török: “Studies of the Growth of an Ascitic Tumour. II. A System to Study the Tumour-Age Dependent Effect of Antitumour Agents”, Eur. J. Cancer, Vol. 14, (1978), pp. 75–82. CrossrefGoogle Scholar

  • [4] A. Kurnatowski and R.G. Willighagen: “Cytochemistry of the NK/Ly Lymphoma”, Nature, Vol. 198, (1961), pp. 1211–1212. http://dx.doi.org/10.1038/1981211a0CrossrefGoogle Scholar

  • [5] M.A. Halma, N.M. Wheelhouse, M.D. Barber, J.J. Powell, K.C. Fearon and J.A. Ross: “Interferon-gamma polymorphisms correlate with duration of survival in pancreatic cancer”, Hum. Immunol., Vol. 65, (2004), pp. 1405–1408. http://dx.doi.org/10.1016/j.humimm.2004.08.184CrossrefGoogle Scholar

  • [6] S. Zawistowski: Technika Histologiczna: Histologia oraz podstawy histopatologii, Panstowy Zaklad Wydawnictwa Lekarskich, Warszawa, 1983 (in Polish). Google Scholar

  • [7] C. Price-Jones: Red Blood Cell Diameters, Oxford Univ. Press, London, 1934. Google Scholar

  • [8] Z. Lojda: “Remarks on biochemical detection of dehydrogenases”, Folia Morphol. (Praha), Vol. 13, (1965), pp. 84–96. Google Scholar

  • [9] O.R. Bessey, O.H. Lowry and M.J. Brock: “A method for the rapid determination of acid phosphatase with five cubic millimeters of serum”, J. Biol. Chem., Vol. 164, (1946), pp. 321–329. Google Scholar

  • [10] N.M. Berezhnaya and V.F. Chekhun: Interleukin System and Cancer, DIA, Kyiv, 2000 (in Russian). Google Scholar

  • [11] N.M Berezhnaya and V.F. Chekhun: Immunology of Malignant Growth, Naukova Dumka, Kyiv, 2005 (in Russian) Google Scholar

  • [12] M.J. Tisdale: “Pathogenesis of Cancer Cachexia”, J. Support Oncol., Vol. 1, (2003), pp. 159–168. Google Scholar

  • [13] S. Rose-John, J. Scheller, G. Elson and S.A. Jones: “Interleukin-6 biology is coordinated by membrane-bound and soluble receptors: role in inflammation and cancer”, J. Leukoc. Biol., Vol. 80, (2006), pp. 227–236. http://dx.doi.org/10.1189/jlb.1105674Google Scholar

  • [14] K. Fujimoto-Ouchi, E. Onuma, M. Shirane, K. Mori and Y. Tanaka: “Capecitabine improves cancer cachexia and normalizes IL-6 and PTHrP levels in mouse cancer cachexia models”, Cancer Chemother. Pharmacol., (2006), Sep 29 (Epub ahead of print). Google Scholar

  • [15] G. Strassmann, M. Fong, J. Kenney and C.O. Jacob: “Evidence for the involvement of interleukin 6 in experimental cancer cachexia”, J. Clin. Invest., Vol. 89, (1992), pp. 1681–1684. http://dx.doi.org/10.1172/JCI115767CrossrefGoogle Scholar

  • [16] J.M. Argiles, S. Busquets and F.J. Lopez-Soriano: “Cytokines in the pathogenesis of cancer cachexia”, Curr. Opin. Clin. Nutr. Metab. Care, Vol. 6, (2003), pp. 401–406. http://dx.doi.org/10.1097/00075197-200307000-00008CrossrefGoogle Scholar

  • [17] M.E. Martignoni, P. Kunze, W. Hildebrandt, B. Kunzli, P. Berberat, T. Giese, O. Kloters, J. Hammer, M.W. Buchler, N.A. Giese and H. Friess: “Role of mononuclear cells and inflammatory cytokines in pancreatic cancer-related cachexia”, Clin. Cancer. Res., Vol. 11, (2005), pp. 5802–5808. http://dx.doi.org/10.1158/1078-0432.CCR-05-0185CrossrefGoogle Scholar

  • [18] S. Tabibzadeh, A. Poubouridis, L. May and P.B. Sehgal: “Interleukin-6 immunoreactivity in human tumors”, Am. J. Pathol., Vol. 135, (1989), pp. 427–433. Google Scholar

  • [19] A. Enomoto, M.C. Rho, A. Fukami, O. Hiraku O, K. Komiyama and M. Hayashi: “Suppression of cancer cachexia by 20S,21-epoxy-resibufogenin-3-acetate-a novel nonpeptide IL-6 receptor antagonist”, Biochem. Biophys. Res. Commun., Vol. 323, (2004), pp. 1096–1102. http://dx.doi.org/10.1016/j.bbrc.2004.08.196Google Scholar

  • [20] M. Trikha, R. Corringham, B. Klein and J.-F. Rossi: “Targeted Anti-Interleukin-6 Monoclonal Antibody Therapy for Cancer: A Review of the Rationale and Clinical Evidence”, Clin. Cancer Res., Vol. 9, (2003), pp. 4653–4665. Google Scholar

  • [21] G. Lelli, M. Montanari, G. Gilli, D. Scapoli, C. Antonietti and D. Scapoli: “Treatment of the cancer anorexia-cachexia syndrome: a critical reappraisal”, J. Chemother., Vol. 15, (2003), pp. 220–225. CrossrefGoogle Scholar

  • [22] Z. Chen, P.S. Malhotra, G.R. Thomas, F.G. Ondrey, D.C. Duffey, C.W. Smith, I. Enamorado, N. T. Yeh, G.S. Kroog, S. Rudy, L. McCullagh, S. Mousa, M. Quezado, L.L. Herscher and C. Van Waes: “Expression of Proinflammatory and Proangiogenic Cytokines in Patients with Head andNeck Cancer”, Clin. Cancer Res., Vol. 5, (1999), pp. 1369–1379. Google Scholar

  • [23] B. Knoefel, K. Nuske, T. Steiner, K. Junker, H. Kosmehi, K. Rebstock, D. Reinhold and U. Junker: “Renal cell carcinoma produce IL-6, IL-10, IL-11 and TGF-beta1 in primary cultures and modulate T lymphocyte blast transformation”, J. Interferon Cytokine Res., Vol. 17, (1997), pp. 95–102. http://dx.doi.org/10.1089/jir.1997.17.95CrossrefGoogle Scholar

  • [24] K.V. Woods, A. El-Naggar, G.L. Clayman and E.A. Grimm: “Variable expression of cytokines in human head and neck squamous carcinoma cell lines and consistent expression in surgical specimens”, Cancer Res., Vol. 58, (1998), pp. 3132–3141 Google Scholar

  • [25] M.G. Alexandrakis, F.H. Passam, E.S. Ganotakis, K. Sfiridaki, I. Xilouri, K. Perisinakis and D.S. Kyriakou: “The clinical and prognostic significance of erythrocyte sedimentation rate ESR, serum interleukin-6 (IL-6) and acute phase protein levels in multiple myeloma”, Clin. Lab. Haematol., Vol. 25, (2000), pp. 41–46. http://dx.doi.org/10.1046/j.1365-2257.2003.00492.xCrossrefGoogle Scholar

  • [26] M.G. Alexandrakis, C.A. Pappa, S. Miyakis, A. Sfiridaki, M. Kafousi, A. Alegakis and E.N. Stathopoulos: “Serum interleukin-17 and its relationship to angiogenic factors in multiple myeloma”, Eur. J. Intern. Med., Vol. 17, (2006), pp. 412–416. http://dx.doi.org/10.1016/j.ejim.2006.02.012CrossrefGoogle Scholar

  • [27] M. Herrmann, H.M. Lorenz, R. Voll, M. Grunke, W. Woith and J.R. Kolden: “A rapid and simple method for the isolation of apoptotic DNA fragments”, Nucl. Acids Res., Vol. 22, (1994), pp. 5506–5507. http://dx.doi.org/10.1093/nar/22.24.5506CrossrefGoogle Scholar

  • [28] R.E. Kast: “Aspirin, TNF-alpha, NFkB, and survival in multiple myeloma: the importance of measuring TNF-alpha”, Inflammopharmacology, Vol. 14, (2006), pp. 256–259. http://dx.doi.org/10.1007/s10787-006-1532-6CrossrefGoogle Scholar

  • [29] T. Kimlinger, M. Kline, S. Kumar, J. Lust, T. Witzig and S.V. Rajkumar: “Differential expression of vascular endothelial growth factors and their receptors in multiple myeloma”, Haematologica, Vol. 91, (2006), pp. 1033–1040. Google Scholar

  • [30] M.I. Koukourakis, D. Papazoglou, A. Giatromanolaki, G. Bougioukas, E. Maltezos and E. Sivridis: “VEGF gene sequence variation defines VEGF gene expression status and angiogenic activity in non-small cell lung cancer”, Lung Cancer, Vol. 43, (2004), pp. 293–298. http://dx.doi.org/10.1016/j.lungcan.2004.04.037CrossrefGoogle Scholar

  • [31] D.R. Senger, S.J. Galli, A.M. Dvorak, C.A. Perruzzi, V.S. Harvey and H.F. Dvorak: “Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid”, Science, Vol. 219, (1983), pp. 983–985. http://dx.doi.org/10.1126/science.6823562CrossrefGoogle Scholar

  • [32] Y. Noguchi, T. Yoshikawa, A. Matsumoto, G. Svaninger and J. Gelin: “Are cytokines possible mediators of cancer cachexia?”, Surg Today, Vol. 26, (1996), pp. 467–475. http://dx.doi.org/10.1007/BF00311551CrossrefGoogle Scholar

  • [33] H. Ishikawa, N. Tsuyama, M. Obata and M. Kawano: “Mitogenic signals initiated via interleukin-6 receptor complexes in cooperation with other transmembrane molecules in myelomas”, J. Clin. Exp. Hematop., Vol. 46, (2006), pp. 55–66. http://dx.doi.org/10.3960/jslrt.46.55CrossrefGoogle Scholar

  • [34] A. Nicolini, A. Carpi and G. Rossi: “Cytokines in breast cancer”, Cytokine Growth Factor Rev., Vol. 17, (2006), pp. 325–337. http://dx.doi.org/10.1016/j.cytogfr.2006.07.002CrossrefGoogle Scholar

  • [35] F. Martin, F. Santolaria, N. Batista, A. Milena, E. Gonzalez-Reimers, M.J. Brito and J. Oramas: “Cytokine levels (IL-6 and IFN-γ), acute phase response and nutritional status as prognostic factors in lung cancer”, Cytokine, Vol. 11, (1999), pp. 80–86. http://dx.doi.org/10.1006/cyto.1998.0398CrossrefGoogle Scholar

  • [36] G. Monti, M.C. Jauraud, I. Monnet, P. Chrdtien, L. Saint-Etienne, L. Zeng, A. Pottier, P. Deviller, P. Galanaud, J. Bignon and D. Emilie: “Intrapleural production of interleukin 6 during mesothelioma and its modulation by gamma-interferon treatment”, Cancer Res., Vol. 54, (1994), pp. 4419–4423. Google Scholar

About the article

Published Online: 2007-03-01

Published in Print: 2007-03-01

Citation Information: Open Life Sciences, Volume 2, Issue 1, Pages 71–86, ISSN (Online) 2391-5412, DOI: https://doi.org/10.2478/s11535-007-0011-4.

Export Citation

© 2007 Versita Warsaw. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

Comments (0)

Please log in or register to comment.
Log in