Stanniocalcin 2 is positively and negatively controlled by 1,25(OH)2D3 and PTH in renal proximal tubular cells
Stanniocalcin 2 is positively and negatively controlled by 1,25(OH)2D3 and PTH in renal proximal tubular cells
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DOI：
10.1677/jme-08-0161
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发表时间：
2009-03-01
影响因子：
3.5
通讯作者：
Takeda, Eiji
中科院分区：
文献类型：
--
作者：
Takei, Yuichiro;Yamamoto, Hironori;Takeda, Eiji
我们以前已经确定了第二个哺乳动物斯钙素（STC 2）在人类和证明，STC 2抑制磷酸盐摄取的负鼠肾近端小管细胞系（负鼠肾（OK）细胞）。然而，在OK细胞中Stc 2基因表达的调节还没有很好地理解。在这项研究中，我们确定了负鼠Stc 2的cDNA序列。负鼠STC 2的氨基酸序列与人STC 2的同源性为78.8%，并且具有保守的推测的N-连接糖基化位点。接下来，我们研究了经典的钙磷调节因子1，25（OH）（2）D-3和PTH在OK细胞中对Stc 2基因表达的调节。在使用亲和纯化的抗STC 2抗体的western blot分析中，STC 2蛋白的分泌被1，25（OH）（2）D-3以剂量依赖的方式刺激。相比之下，PTH抑制1，25（OH）（2）D-3诱导的STC 2蛋白分泌。实时荧光定量PCR分析显示，1，25（OH）（2）D-3以剂量和时间依赖的方式增加Stc 2 mRNA的表达。此外，放线菌素D，RNA合成抑制剂，阻止1，25（OH）（2）D-3对Stc 2基因表达的影响。另一方面，PTH和佛波醇12，13-肉豆蔻乙酸酯，一种特异性PKC激活剂，但不是8-溴环AMP，一种特异性PKA激活剂，降低Stc 2的mRNA水平。此外，一种特异性的PKC抑制剂Go 6976可消除PTH对Stc 2 mRNA表达的下调。此外，我们证明，肾Stc 2 mRNA表达增加1，25（OH）（2）D-3和PTH在体内降低。结果提示，STC 2在肾小管上皮细胞中受1，25（OH）（2）D-3和PTH的正性和负性调控。分子内分泌学杂志（2009）42，261-268
We have previously identified a second mammalian stanniocalcin (STC2) in humans and demonstrated that STC2 inhibits phosphate uptake in an opossum renal proximal tubular cell line (opossum kidney (OK) cells). However, the regulation of Stc2 gene expression in OK cells is not well understood. In this study, we identified the opossum Stc2 cDNA sequence. The opossum STC2 amino acid sequence had 78.8% homology with human STC2, and has a conserved putative N-linked glycosylation site. Next, we investigated the regulation of Stc2 gene expression by the classical calcium and phosphate-regulating factors 1,25(OH)(2)D-3 and PTH in OK cells. In western blot analysis using affinity-purified anti-STC2 antibody, the secretion of STC2 protein was stimulated by 1,25(OH)(2)D-3 in a dose-dependent manner. By contrast, PTH suppressed the induction of STC2 protein secretion by 1,25(OH)(2)D-3. Real-time PCR analysis revealed that Stc2 mRNA expression was increased by 1,25(OH)(2)D-3 in a dose- and time-dependent manner. In addition, actinomycin D, an RNA synthesis inhibitor, prevented the effects of 1,25(OH)(2)D-3 on Stc2 gene expression. On the other hand, PTH and phorbol 12,13-myristic acetate, a specific PKC activator, but not 8-bromo-cyclic AMP, a specific PKA activator, reduced the mRNA levels of Stc2. In addition, Go6976, a specific PKC inhibitor, abolished the downregulation of Stc2 mRNA expression by PTH. Furthermore, we demonstrated that the renal Stc2 mRNA expression was increased by 1,25(OH)(2)D-3 and decreased by PTH in vivo. These results suggest that STC2 is positively and negatively controlled by 1,25(OH)(2)D-3 and PTH in renal proximal tubular cells. Journal of Molecular Endocrinology (2009) 42, 261-268