Changes in cytokine production and morphology of murine lymphoma NK/Ly cells in course of tumor development |
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Authors: | Rostyslav R Panchuk Natalia M Boiko Maxim D Lootsik Rostyslav S Stoika |
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Institution: | (1) Institute of Cell Biology, National Academy of Sciences of Ukraine, 79005 Lviv, Ukraine |
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Abstract: | 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. |
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Keywords: | Murine lymphoma NK/Ly cytokines tumor ageing |
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