The Pituitary Center, Department of Medicine, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA.
Endocrinology. 2013 Jul;154(7):2399-409. doi: 10.1210/en.2013-1132. Epub 2013 May 21.
Somatostatin signals predominantly through somatostatin receptor (SSTR) subtype 2 to attenuate GH release. However, the independent role of the receptor in regulating GH synthesis is unclear. Because we had previously demonstrated constitutive SSTR2 activity in mouse corticotrophs, we now analyzed GH regulation in rat pituitary somatotroph (GC) tumor cells, which express SSTR2 exclusively and are devoid of endogenous somatostatin ligand. We demonstrate that moderately stable SSTR2 overexpression (GpSSTR2(WT) cells) was associated with decreased GH promoter activity, GH mRNA, and hormone levels compared with those of control transfectants (GpCon cells). In contrast, levels of GH mRNA and peptide and GH promoter activity were unchanged in GpSSTR2(DRY) stable transfectants moderately expressing DRY motif mutated SSTR2 (R140A). GpSSTR(2DRY) did not exhibit an enhanced octreotide response as did GpSSTR2(WT) cells; however, both SSTR2(WT)-enhanced yellow fluorescent protein (eYFP) and SSTR2(DRY)-eYFP internalized on octreotide treatment. Suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, increased GH synthesis in wild-type GC cells and primary pituitary cultures. GpSSTR2(WT) cells induced GH synthesis more strongly on SAHA treatment, evident by both higher GH peptide and mRNA levels compared with the moderate but similar GH increase observed in GpCon and GpSSTR2(DRY) cells. In vivo SAHA also increased GH release from GpSSTR2(WT) but not from control xenografts. Endogenous rat GH promoter chromatin immunoprecipitation showed decreased baseline acetylation of the GH promoter with exacerbated acetylation after SAHA treatment in GpSSTR2(WT) compared with that of either GpSSTR(2DRY) or control cells, the latter 2 transfectants exhibiting similar GH promoter acetylation levels. In conclusion, modestly increased SSTR2 expression constitutively decreases GH synthesis, an effect partially mediated by GH promoter histone deacetylation.
生长抑素主要通过生长抑素受体 (SSTR) 亚型 2 发出信号,从而减弱 GH 的释放。然而,受体在调节 GH 合成中的独立作用尚不清楚。由于我们之前已经证明了小鼠脑垂体促皮质素细胞中的 SSTR2 存在组成型活性,因此我们现在分析了仅表达 SSTR2 且缺乏内源性生长抑素配体的大鼠垂体生长激素瘤 (GC) 细胞中的 GH 调节。我们证明,与对照转染体 (GpCon 细胞) 相比,中度稳定的 SSTR2 过表达 (GpSSTR2(WT) 细胞) 与 GH 启动子活性、GH mRNA 和激素水平降低相关。相比之下,中度表达 DRY 基序突变的 SSTR2 (R140A) 的 GpSSTR2(DRY) 稳定转染体的 GH mRNA 和肽水平以及 GH 启动子活性保持不变。GpSSTR(2DRY) 没有像 GpSSTR2(WT) 细胞那样表现出增强的奥曲肽反应;然而,SSTR2(WT)-增强型黄色荧光蛋白 (eYFP) 和 SSTR2(DRY)-eYFP 都在奥曲肽处理时内化。琥珀酰亚胺基羟肟酸 (SAHA),一种组蛋白去乙酰化酶抑制剂,增加了野生型 GC 细胞和原代垂体培养物中的 GH 合成。GpSSTR2(WT) 细胞在用 SAHA 处理时诱导 GH 合成的作用更强,这表现为与 GpCon 和 GpSSTR2(DRY) 细胞中度但相似的 GH 增加相比,GH 肽和 mRNA 水平均更高。体内 SAHA 也增加了 GpSSTR2(WT) 而非对照异种移植物中的 GH 释放。内源性大鼠 GH 启动子染色质免疫沉淀显示,与 GpSSTR(2DRY) 或对照细胞相比,SAHA 处理后 GH 启动子的基础乙酰化水平降低,后者 2 种转染体的 GH 启动子乙酰化水平相似。总之,适度增加的 SSTR2 表达会持续降低 GH 合成,这种作用部分通过 GH 启动子组蛋白去乙酰化来介导。