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Effect of TNF-α on Raji cells at different cellular levels estimated by various methods

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Abstract

Tumor necrosis factor (TNF)-α, a pleiotropic cytokine, has been shown to induce diverse and opposite effects on lymphoid malignancy depending on TNF receptor system expression. Based on this, we investigated its in vitro dose- and time-related effect on the malignant B-cell line Raji, derived from Burkitt lymphoma patients, at different intracellular levels. The membrane alteration was estimated by lactate dehydrogenase (LDH) release and by flow cytometry; intracellular metabolic energy by determination of the total intracellular LDH activity; total cytosole protein mass by sulforhodamine B assay; and cell growth by incorporation of [3H]thymidine into DNA. Significant increase of LDH through cell membrane alteration was accompanied by decrease of intracellular metabolized energy and total protein mass. TNF-α at lower concentrations (125 and 250 pg/ml) significantly induced cell proliferation in comparison with 1,000 pg/ml of TNF-α, which induced more cell death. TNF-α induced maximal apoptosis rate up to 30% after 24 h, showing more effects for a necrotic form of cell death. Here we reported opposite and diverse effects of TNF-α at different intracellular levels in Raji cells, when applied in different assays, showing characteristics for every cellular compartment.

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Acknowledgements

This work was partly supported by a grant from the Serbian Ministry of Science, Technology and Development. We thank Dr. Haral Wajant, Germany, for the TNF-α.

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Authors and Affiliations

  1. School of Medicine, University of Kragujevac, P.O. BOX 124, 34 000, Kragujevac, Serbia and Montenegro

    Vladimir Jurisic

  2. Institute of Oncology, Sremska Kamenica, Serbia and Montenegro

    Gordana Bogdanovic, Vesna Kojic, Dimitar Jakimov & Tatjana Srdic

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  1. Vladimir Jurisic
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  2. Gordana Bogdanovic
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  3. Vesna Kojic
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  4. Dimitar Jakimov
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Correspondence to Vladimir Jurisic.

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Jurisic, V., Bogdanovic, G., Kojic, V. et al. Effect of TNF-α on Raji cells at different cellular levels estimated by various methods. Ann Hematol 85, 86–94 (2006). https://doi.org/10.1007/s00277-005-0010-3

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