TSH stimulation of human thyroglobulin and thyroid peroxidase gene transcription is partially dependent on internalization.
TSH stimulation of human thyroglobulin and thyroid peroxidase gene transcription is partially dependent on internalization.
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DOI：
10.1016/j.cellsig.2021.110212
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发表时间：
2022-03
影响因子：
4.8
通讯作者：
Gershengorn MC
中科院分区：
文献类型：
--
作者：
Jang D;Eliseeva E;Klubo-Gwiezdzinska J;Neumann S;Gershengorn MC
TSH 受体 (TSHR) 是人类甲状腺细胞中甲状腺激素生物合成的主要调节因子，通过调节包括甲状腺球蛋白 (TG) 和甲状腺过氧化物酶 (TPO) 在内的许多基因的转录。直到最近，人们还认为 TSHR 仅在细胞表面启动信号转导途径，而内化主要涉及 TSHR 脱敏和下调。主要在小鼠细胞中进行的研究表明，TSHR 内化也调节细胞内位点的基因转录。然而，对于参与人类甲状腺细胞中甲状腺激素生物合成的基因，这一点尚未得到证实。我们在原代培养中使用人类甲状腺细胞。在这些细胞中，TSH 基因表达的剂量反应是双相的，低剂量上调基因表达，高剂量降低基因表达。我们使用两种方法来抑制内化。首先，我们使用了 dynamins、dynasore 和 dyngo-4a 抑制剂。 dynasore 或 dyngo-4a 预处理显着抑制 TG 和 TPO mRNA 的 TSH 上调以及 TG 分泌。在第二步中，我们使用了动力蛋白 2 的敲低，动力蛋白 2 是人类甲状腺细胞中最丰富的动力蛋白。我们发现，dynamin 2 敲低可抑制 TSHR 内化，并降低 TSH 刺激的 TG 和 TPO mRNA 和蛋白质水平。最后，我们发现，当内化受到抑制时，细胞核中激活转录因子磷酸化 cAMP 反应元件结合蛋白 (pCREB) 的水平降低了 68%。我们的结论是，人类甲状腺细胞中甲状腺激素合成相关基因的上调部分依赖于导致激活转录因子核定位的内化。
The TSH receptor (TSHR) is the major regulator of thyroid hormone biosynthesis in human thyrocytes by regulating the transcription of a number of genes including thyroglobulin (TG) and thyroperoxidase (TPO). Until recently, it was thought that TSHR initiated signal transduction pathways only at the cell-surface and that internalization was primarily involved in TSHR desensitization and downregulation. Studies primarily in mouse cells showed that TSHR internalization regulates gene transcription at an intracellular site also. However, this has not been shown for genes involved in thyroid hormone biosynthesis in human thyrocytes. We used human thyrocytes in primary culture. In these cells, the dose-response to TSH for gene expression is biphasic with low doses upregulating gene expression and higher doses decreasing gene expression. We used two approaches to inhibit internalization. In the first, we used inhibitors of dynamins, dynasore and dyngo-4a. Pretreatment with dynasore or dyngo-4a markedly inhibited TSH upregulation of TG and TPO mRNAs, as well as TG secretion. In the second, we used knockdown of dynamin 2, which is the most abundant dynamin in human thyrocytes. We showed that dynamin 2 knockdown inhibited TSHR internalization and decreased the TSH-stimulated levels of TG and TPO mRNAs and proteins. Lastly, we showed that the level of the activatory transcription factor phosphorylated cAMP response element binding protein (pCREB) in the cell nuclei was reduced by 68% when internalization was inhibited. We conclude that upregulation of genes involved in thyroid hormone synthesis in human thyrocytes is, in part, dependent on internalization leading to nuclear localization of an activated transcription factor(s).
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DOI：
10.1007/978-1-4939-8600-2_1
发表时间：
2018
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Morgan SJ;Neumann S;Gershengorn MC
通讯作者：
Gershengorn MC
影响因子：
2.1
作者：
ABADIA, J
通讯作者：
ABADIA, J
影响因子：
--
作者：
Baratti-Elbaz, C;Ghinea, N;Milgrom, E
通讯作者：
Milgrom, E
影响因子：
4.8
作者：
Neumann, Susanne;Geras-Raaka, Elizabeth;Gershengorn, Marvin C.
通讯作者：
Gershengorn, Marvin C.
影响因子：
9.8
作者：
Calebiro D;Nikolaev VO;Gagliani MC;de Filippis T;Dees C;Tacchetti C;Persani L;Lohse MJ
通讯作者：
Lohse MJ