Taki Katsumi, Kogai Takahiko, Kanamoto Yoko, Hershman Jerome M, Brent Gregory A
Endocrinology Division, Veterans Affairs Greater Los Angeles Healthcare System and Department of Medicine, University of California-Los Angeles School of Medicine, Los Angeles, California 90073, USA.
Mol Endocrinol. 2002 Oct;16(10):2266-82. doi: 10.1210/me.2002-0109.
The sodium/iodide symporter (NIS) gene is highly expressed in the thyroid gland and is important for the diagnosis and radioiodide therapy of differentiated thyroid cancers. We investigated a human NIS (hNIS) gene 5'-far-upstream enhancer (hNUE) (-9847 to -8968). The hNUE is TSH responsive in both FRTL-5 cells and primary normal thyroid cells, but not in human papillary thyroid cancer cells (BHP cells). The hNUE enhanced expression of the basal hNIS promoter 15-fold and required both a Pax-8 binding site and a cAMP response element (CRE)-like sequence for full activity. The hNUE activated transcription in a thyroid-selective and cAMP-dependent manner, mediated by both protein kinase A (PKA)-dependent and PKA-independent pathways. Pax-8 and two CRE-like sequence binding proteins bind to the hNUE. Supershift binding assay indicated that one of the CRE-like sequence binding protein(s) was CRE-binding protein-1, activation transcription factor-1, and/or CRE modulator, and the other was an unknown factor(s) that is absent in BHP 2-7 cells. A far-upstream enhancer is important for hNIS regulation in the thyroid. Deficient CRE-like sequence binding protein(s) that bind to the hNUE in normal thyroid cells may be responsible for reduced NIS gene expression in some thyroid carcinomas.
钠/碘同向转运体(NIS)基因在甲状腺中高度表达,对分化型甲状腺癌的诊断和放射性碘治疗具有重要意义。我们研究了人NIS(hNIS)基因5'端远上游增强子(hNUE)(-9847至-8968)。hNUE在FRTL-5细胞和原代正常甲状腺细胞中对促甲状腺激素(TSH)有反应,但在人甲状腺乳头状癌细胞(BHP细胞)中无反应。hNUE使基础hNIS启动子的表达增强了15倍,且其充分发挥活性需要一个Paired box 8(Pax-8)结合位点和一个类似环磷酸腺苷(cAMP)反应元件(CRE)的序列。hNUE以甲状腺选择性和cAMP依赖性方式激活转录,由蛋白激酶A(PKA)依赖性和PKA非依赖性途径介导。Pax-8和两种类似CRE序列的结合蛋白与hNUE结合。超迁移结合试验表明,其中一种类似CRE序列的结合蛋白是CRE结合蛋白-1、激活转录因子-1和/或CRE调节剂,另一种是BHP 2-7细胞中不存在的未知因子。远上游增强子对甲状腺中hNIS的调控很重要。正常甲状腺细胞中与hNUE结合的类似CRE序列的结合蛋白缺陷可能是某些甲状腺癌中NIS基因表达降低的原因。