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Retinoic acid receptor β2 re-expression and growth inhibition in thyroid carcinoma cell lines after 5-aza-2′-deoxycytidine treatment

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Abstract

The treatment of both undifferentiated and de-differentiated thyroid tumors, which are unresponsive to radioiodine, represents one of the biggest challenges for thyroidologists. The aim of the present study was to investigate in vitro the methylation status of retinoic acid receptors (RAR)β2 promoter and the effect of the demethylating agent 5-aza-2′-deoxycytidine (5-Aza-CdR) on 5 human thyroid cancer cell lines. The methylation status of RARβ2 promoter was analyzed by methylation-specific PCR. The effect of 5-Aza-CdR on cell growth and apoptosis was evaluated by cell counting, enzyme-linked immunosorbent assay tests and fluorescence-activated cell sorting analysis, while the effect on the expression of RAR and thyroid-specific genes was measured by qualitative and quantitative RT-PCR. Methylation of RARβ2 promoter was present only in ARO cells. 5-Aza-CdR determined growth inhibition in all cell lines, probably due to apoptosis in WRO, NPA, and ARO cells, and to inhibition of DNA synthesis in TT cells. Treatment with 5-Aza-CdR induced the expression of RARβ mRNA in ARO and FRO cells, a slight increase of the expression of Tg, TPO and thyroid trancription factor 1 (TTF-1) mRNA and the new expression of low levels of NIS in TT cells. A significant increase of TTF-1 mRNA in FRO cells was also observed. In this study we demonstrated that RARβ2 promoter was methylated in ARO cell line. However, the 5-Aza-CdR treatment induced RARβ mRNA expression not only in ARO but also in FRO and TT cell lines, whose RARβ2 promoter was unmethylated. A significant reduction of cell growth, but not cell re-differentiation, was also observed after 5-Aza-CdR treatment.

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Authors and Affiliations

  1. Department of Endocrinology and Metabolism, University of Pisa, Via Paradisa no 2, 56124, Pisa, Italy

    F. Y. Miasaki, A. Vivaldi, R. Ciampi, L. Agate, A. Pinchera & R. Elisei MD

  2. Department of Oncology, Division of Pathology I, Italy

    P. Collecchi & A. Capodanno

  3. AMBISEN Center, High Technology Center for the Study of the Environmental Damage of the Endocrine and Nervous Systems, University of Pisa, Pisa, Italy

    A. Pinchera

Authors
  1. F. Y. Miasaki
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  2. A. Vivaldi
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  3. R. Ciampi
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  4. L. Agate
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  5. P. Collecchi
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  8. R. Elisei MD
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Correspondence to R. Elisei MD.

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Miasaki, F.Y., Vivaldi, A., Ciampi, R. et al. Retinoic acid receptor β2 re-expression and growth inhibition in thyroid carcinoma cell lines after 5-aza-2′-deoxycytidine treatment. J Endocrinol Invest 31, 724–730 (2008). https://doi.org/10.1007/BF03346422

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