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Induction of apoptosis in T lymphoma cells by long-term treatment with thyroxine involves PKCζ nitration by nitric oxide synthase

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Abstract

Thyroid hormones are important regulators of cell physiology, inducing cell proliferation, differentiation or apoptosis, depending on the cell type. Thyroid hormones induce proliferation in short-term T lymphocyte cultures. In this study, we assessed the effect of long-term thyroxine (T4) treatment on the balance of proliferation and apoptosis and the intermediate participants in T lymphoma cells. Treatment with T4 affected this balance from the fifth day of culture, inhibiting proliferation in a time-dependent manner. This effect was associated with apoptosis induction, as characterized through nuclear morphological changes, DNA fragmentation, and Annexin V-FITC/Propidium Iodide co-staining. In addition, increased iNOS gene and protein levels, and enzyme activity were observed. The generation of reactive oxygen species, depolarization of the mitochondrial membrane, and a reduction in glutathione levels were also observed. The imbalance between oxidants and antioxidants species is typically associated with the nitration of proteins, including PKCζ, an isoenzyme essential for lymphoma cell division and survival. Consistently, evidence of PKCζ nitration via proteasome degradation was also observed in this study. Taken together, these results suggest that the long-term culture of T lymphoma cells with T4 induces apoptosis through the increased production of oxidative species resulting from both augmented iNOS activity and the loss of mitochondrial function. These species induce the nitration of proteins involved in cell viability, promoting proteasome degradation. Furthermore, we discuss the impact of these results on the modulation of T lymphoma growth and the thyroid status in vivo.

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Acknowledgments

The authors wish to thank Mrs María Rosa Gonzalez Murano for her excellent technical assistance. This work was supported by Grants from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), PIP-CONICET N° 00275, from Agencia Nacional para la Promoción Científica y Técnica, PICT 2008 N° 1858 and from University of Buenos Aires, UBACYT N° 20020100100291.

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Authors and Affiliations

  1. Instituto de Investigaciones Biomédicas, Facultad de Ciencias Médicas, Pontificia Universidad Católica Argentina (UCA), CONICET, Alicia M. de Justo 1600 3° piso, CABA, Buenos Aires, Argentina

    M. L. Barreiro Arcos, H. A. Sterle, E. Valli, M. F. Cayrol, A. J. Klecha, M. A. Paulazo, M. C. Diaz Flaqué & G. A. Cremaschi

  2. Instituto de Investigación en Biomedicina de Buenos Aires (CONICET), Buenos Aires, Argentina

    C. Vercelli

  3. Centro de Estudios Farmacológicos y Botánicos (CEFYBO), CONICET, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    A. M. Franchi

  4. Laboratorio de Radioisótopos, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina

    A. J. Klecha & G. A. Cremaschi

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  1. M. L. Barreiro Arcos
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  2. H. A. Sterle
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  3. C. Vercelli
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  4. E. Valli
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  5. M. F. Cayrol
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  9. A. M. Franchi
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  10. G. A. Cremaschi
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Barreiro Arcos, M.L., Sterle, H.A., Vercelli, C. et al. Induction of apoptosis in T lymphoma cells by long-term treatment with thyroxine involves PKCζ nitration by nitric oxide synthase. Apoptosis 18, 1376–1390 (2013). https://doi.org/10.1007/s10495-013-0869-8

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