PCP4: a regulator of aldosterone synthesis in human adrenocortical tissues.
PCP4: a regulator of aldosterone synthesis in human adrenocortical tissues.
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DOI：
10.1530/jme-13-0248
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发表时间：
2014-04
影响因子：
3.5
通讯作者：
Sasano H
中科院分区：
文献类型：
--
作者：
Felizola SJ;Nakamura Y;Ono Y;Kitamura K;Kikuchi K;Onodera Y;Ise K;Takase K;Sugawara A;Hattangady N;Rainey WE;Satoh F;Sasano H
浦肯野细胞蛋白4（Purkinje cell protein 4，PCP 4）是一种钙调素（CaM）结合蛋白，可加速钙离子与CaM的结合和解离。它以前曾在醛固酮腺瘤（阿帕）中检测到，但其在肾上腺皮质组织中的表达和功能的细节仍然未知。因此，我们对以下组织中的PCP 4进行了免疫组织化学分析：正常肾上腺（NA; n=15）、阿帕（n= 15）、皮质醇生成腺瘤（CPA; n=15）和特发性醛固酮增多症病例（IHA; n=5）。阿帕样本（n=45）还接受了PCP 4、CYP 11B 1和CYP 11B 2的定量RT-PCR（qPCR），以及KCNJ 5突变的DNA测序。在ELISA分析之后，还在H295 R肾上腺皮质癌细胞中进行了使用PCP 4 siRNA的瞬时转染分析，并且在PCP 4载体转染后还进行了CYP 11B 2荧光素酶测定，以研究PCP 4蛋白表达的调节。在我们的研究结果中，PCP 4免疫反应主要在阿帕和NA和IHA的肾小球（ZG）中检测到。阿帕患者PCP 4和CYP 11 B2的mRNA表达水平显著相关（P<0.0001），KCNJ 5突变患者PCP 4 mRNA表达水平显著高于野生型患者（P=0.005）。PCP 4载体转染后，CYP 11B 2荧光素酶报告基因活性显著高于血管紧张素II存在下的对照组。敲低PCP 4导致CYP 11B 2 mRNA水平（P=0.012）和醛固酮生成（P=0.011）显著降低。我们的研究结果表明，PCP 4是正常的，增生和肿瘤的人肾上腺皮质细胞的醛固酮生产的调节器。
Purkinje cell protein 4 (PCP4) is a calmodulin (CaM) binding protein that accelerates calcium association and dissociation with CaM. It has been previously detected in aldosterone-producing adenomas (APA) but details on its expression and function in adrenocortical tissues have remained unknown. Therefore, we performed the immunohistochemical analysis of PCP4 in the following tissues: normal adrenal (NA; n=15), APA (n=15), cortisol producing adenomas (CPA; n=15) and idiopathic hyperaldosteronism cases (IHA; n=5). APA samples (n=45) were also submitted to quantitative RT-PCR (qPCR) of PCP4, CYP11B1, and CYP11B2, as well as DNA sequencing for KCNJ5 mutations. Transient transfection analysis using PCP4 siRNA was also performed in H295R adrenocortical carcinoma cells, following ELISA analysis, and CYP11B2 luciferase assays were also performed after PCP4 vector transfection in order to study the regulation of PCP4 protein expression. In our findings, PCP4 immunoreactivity was predominantly detected in APA and in the zona glomerulosa (ZG) of NA and IHA. In APA, the mRNA levels of PCP4 were significantly correlated with those of CYP11B2 (P<0.0001) and were significantly higher in cases with KCNJ5 mutation than wild-type (P=0.005). Following PCP4 vector transfection, CYP11B2 luciferase reporter activity was significantly higher than controls in the presence of angiotensin-II. Knockdown of PCP4 resulted in a significant decrease in CYP11B2 mRNA levels (P=0.012) and aldosterone production (P=0.011). Our results indicate that PCP4 is a regulator of aldosterone production in normal, hyperplastic and neoplastic human adrenocortical cells.