Pituitary adenylate cyclase-activating polypeptide inhibits transforming growth factor-beta1-induced apoptosis in a human pituitary adenoma cell line.
Pituitary adenylate cyclase-activating polypeptide inhibits transforming growth factor-beta1-induced apoptosis in a human pituitary adenoma cell line.
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垂体腺苷酸环化酶激活多肽抑制人垂体腺瘤细胞系中转化生长因子-β1 诱导的细胞凋亡。
DOI：
10.1016/s0002-9440(10)65509-5
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Lloyd,RV
中科院分区：
文献类型：
--
作者：
Oka,H;Jin,L;Kulig,E;Scheithauer,BW;Lloyd,RV
垂体腺苷酸环化酶激活多肽（PACAP）最初是从下丘脑组织中分离出来的，基于其刺激培养的垂体前叶细胞产生cAMP的能力。最近的研究表明，PACAP在脑细胞凋亡中的功能作用。然而，PACAP在调节人垂体腺瘤细胞凋亡中的作用以前没有被研究过。对表达所有三种主要PACAP受体的培养的人垂体腺瘤细胞系HP 75的分析表明，PACAP-38和PACAP-27均抑制TGF-β1诱导的凋亡。用PACAP受体拮抗剂PACAP 6-38（PACAP I型受体拮抗剂）和（p-chloro-d-Phe 6，Leu 17）-VIP（PACAP II型受体拮抗剂）处理可阻断PACAP-38对转化生长因子-β1（TGF-β1）诱导的细胞凋亡的抑制作用，证实了PACAP作用的特异性。佛司可林（forskolin）处理也能抑制TGF-β1诱导的细胞凋亡，而佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）处理则无此作用。通过Western印迹分析，TGF-β1处理与丝裂原活化蛋白激酶（MAP激酶）的增加相关，但PACAP抑制TGF-β1诱导的细胞凋亡与MAP激酶的活化无关。细胞周期细胞周期蛋白依赖性激酶抑制剂p27的免疫细胞化学分析表明，用TGF-β1、毛喉素、PMA和PACAP处理可增加培养的HP 75细胞中p27的表达。这些结果表明，PACAP是HP 75人垂体腺瘤细胞系中TGF-β1诱导的凋亡的高度特异性抑制剂，并且PACAP、TGF-β1、forskolin和PMA都刺激HP 75人垂体腺瘤细胞系中TGF-β调节的细胞周期蛋白p27的表达。HP 75细胞系可作为研究人垂体细胞凋亡调控的模型。
Pituitary adenylate cyclase-activating polypeptide (PACAP) was originally isolated from hypothalamic tissues based on its ability to stimulate cAMP production in cultured anterior pituitary cells. Recent studies have suggested a functional role for PACAP in the apoptosis of brain cells. However, the role of PACAP in regulating apoptosis in human pituitary adenomas has not previously been examined. Analysis of the cultured human pituitary adenoma cell line HP75, which expresses all three major PACAP receptors, showed that both PACAP-38 and PACAP-27 inhibited TGF-β1-induced apoptosis. Treatment with the PACAP receptor antagonists PACAP 6–38 (PACAP type I receptor antagonist) and (p-chloro-d-Phe6, Leu17)-VIP (PACAP type II receptor antagonist) blocked the effects of PACAP-38 on the inhibition of transforming growth factor-β1 (TGF-β1)-induced apoptosis, confirming the specificity of the role of PACAP. Treatment with forskolin but not phorbol 12-myristate 13-actetate (PMA) also inhibited TGF-β1-induced apoptosis. TGF-β1 treatment was associated with an increase in mitogen-activated protein kinase (MAP kinase) when analyzed by Western blotting, but PACAP inhibition of TGF-β1-induced apoptosis was not associated with activation of MAP kinase. Immunocytochemical analysis of the cell cycle cyclin-dependent kinase inhibitor p27 showed that treatment with TGF-β1, forskolin, PMA, and PACAP increased p27 expression in cultured HP75 cells. These results indicate that PACAP is a highly specific inhibitor of TGF-β1-induced apoptosis in the HP75 human pituitary adenoma cell line and that PACAP, TGF-β1, forskolin, and PMA all stimulate expression of the TGF-β-regulated cell cycle protein p27 in the HP75 human pituitary adenoma cell line. The HP75 cell line can be used as a model to study the regulation of apoptosis in human pituitary cells.
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