Transient modulation of cytoplasmic and nuclear retinoid receptors expression in differentiating human teratocarcinoma NT2 cells

Laboratory of Molecular Biology, National Cancer Research Institute (IST), Genova, Italy.
Journal of Neurochemistry (Impact Factor: 4.28). 02/2003; 84(1):94-104. DOI: 10.1046/j.1471-4159.2003.01501.x
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ABSTRACT Human embryonal carcinoma Ntera2/D1 (NT2) cells treated with retinoic acid (RA) differentiate into several cell types including post-mitotic neurons. In this study we asked if RA-induced differentiation alters the expression of RA and retinol (ROL) binding proteins. The regulation of the intracellular carrier proteins for ROL and RA, cellular retinol binding protein I (CRBP-I), and cellular retinoic acid binding protein I and II (CRABP-I, CRABP-II) were studied along with the nuclear RA receptors RARalpha, RARbeta and RARgamma2. PCR analysis of total mRNA from RA-treated cells showed a biphasic early induction of CRBP-I, CRABP-II, and RARgamma2 genes. The immediate early gene Krox-24, a zinc finger transcription factor which is up-regulated during neuronal differentiation, was also induced, but after 1 week of treatment. The induction of CRBP-I protein synthesis in differentiating NT2 cells was confirmed by western blotting and immunofluorescence experiments. Conversely, the synthetic retinoid N-(4-hydroxyphenyl)retinamide, which induces cell death, but not differentiation in different tumour cell types, did not produce the same modulation on gene expression in NT2 cells. These data suggest that the RA-specific induction of CRBP-I and CRABP-II could be an early event in the process leading to neuronal differentiation of NT2 cells.

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Available from: Francesca Isabella Tosetti, Sep 28, 2015
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    • "RT-PCR was performed as described previously [4] [22] [25]. The synthesized cDNA was amplified using specific primer sets for b-actin or cellular retinol binding protein I (CRBP-I), as described previously [26] [27]. The sequence of the primer set for ZNF267 was 5 0 -CCTTTCGCTGTAGTTCATAC-3 0 and 5 0 -CCGATGCACAG AAAGACCT-3 0 , the expected length of the amplified product was 538 bp. "
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