Gi-protein inhibitor, guanosine 5'-O-(2-thiodiphosphate), induces senescence-associated beta-galactosidase positive cell formation through CREB phosphorylation.

Moon EY; Kim HS; Im YS

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Gi-protein inhibitor, guanosine 5'-O-(2-thiodiphosphate), induces senescence-associated beta-galactosidase positive cell formation through CREB phosphorylation.

机译：Gi蛋白抑制剂鸟苷5'-O-（2-硫代二磷酸）通过CREB磷酸化诱导衰老相关的β-半乳糖苷酶阳性细胞。

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AIMS: We evaluated Gi-protein inhibitor, guanosine 5'-O-(2-thiodiphosphate)(GOT)-induced senescence-associated(SA)-beta-galactosidase(Gal) positive cell formation to determine if it occurred through phosphorylation of cyclic AMP-dependent response element binding protein (CREB). MAIN METHODS: IMR-90 human lung fibroblast cells were used. SA-beta-Gal positive cells and senescence-associated heterochromatic foci (SAHF) were determined by assessing blue color formation of substrate, X-gal inside cells and DAPI staining, respectively. Cell cycle and hypodiploid cell formation were assessed by flow cytometry analysis. CREB phosphorylation and molecular changes were analyzed by western blot. KEY FINDINGS: GOT treatment led to SA-beta-Gal positive cell formation and SAHF. CREB phosphorylation increased in response to GOT treatment but then decreased over 24h. SA-beta-Gal positive cell formation increased in response to transient transfection of pS6-RSV-CREB and no changes were detected following CREB knockdown with CREB-siRNA. In addition, CREB phosphorylation was delayed by treatment with the anti-cellular senescence agents, clitocybins which also reduced the number of SA-beta-Gal positive cells. Collectively, our data showed that GOT-induced CREB phosphorylation initiated SA-beta-Gal positive cell formation after which decreased in SA-beta-Gal positive cells. SIGNIFICANCE: These findings suggest for the first time that CREB phosphorylation by GOT could induce cellular senescence as judged by SA-beta-Gal positive cell formation.

机译：目的：我们评估了Gi蛋白抑制剂鸟苷5'-O-（2-硫代二磷酸）（GOT）诱导的衰老相关（SA）-β-半乳糖苷酶（Gal）阳性细胞形成，以确定其是否通过环磷酸化而发生AMP依赖性反应元件结合蛋白（CREB）。主要方法：使用IMR-90人肺成纤维细胞。通过分别评估底物的蓝色形成，细胞内的X-gal和DAPI染色来确定SA-β-Gal阳性细胞和衰老相关的异色病灶（SAHF）。通过流式细胞术分析评估细胞周期和二倍体细胞形成。通过蛋白质印迹分析CREB的磷酸化和分子变化。主要发现：GOT治疗导致SA-β-Gal阳性细胞形成和SAHF。响应GOT处理，CREB磷酸化增加，但在24小时后降低。响应pS6-RSV-CREB的瞬时转染，SA-β-Gal阳性细胞的形成增加，在用CREB-siRNA敲除CREB后未检测到变化。此外，CREB的磷酸化被抗细胞衰老剂-clitocybins的治疗所延迟，这也减少了SA-β-Gal阳性细胞的数量。总的来说，我们的数据显示GOT诱导的CREB磷酸化启动了SA-β-Gal阳性细胞的形成，此后在SA-β-Gal阳性细胞中减少。意义：这些发现首次表明，根据SA-β-Gal阳性细胞形成判断，GOT的CREB磷酸化可诱导细胞衰老。

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《Life sciences》 |2010年第18期|共8页
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Moon EY; Kim HS; Im YS;
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中图分类生命的起源;生命科学总论;
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1. Gi-protein inhibitor, guanosine 5'-O-(2-thiodiphosphate), induces senescence-associated beta-galactosidase positive cell formation through CREB phosphorylation. [J] . Moon EY, Kim HS, Im YS Life sciences . 2010,第17a18期

机译：Gi蛋白抑制剂鸟苷5'-O-（2-硫代二磷酸）通过CREB磷酸化诱导衰老相关的β-半乳糖苷酶阳性细胞。
2. Involvement of K~+ channel permeability changes in the L-NAME and indomethacin resistant part of adenosine-5'-O- (2-thiodiphosphate)-induced relaxation of pancreatic vascular bed [J] . D.Hillaire-Buys, J.Chapal, N.Linck British Journal of Pharmacology . 1998,第1期

机译：K〜+通道通透性变化参与腺苷5'-O-（2-硫代二磷酸）诱导的胰血管床松弛的L-NAME和吲哚美辛耐药部分
3. Study of the mechanisms involved in adenosine-5′-O-(2-thiodiphosphate) induced relaxation of rat thoracic aorta and pancreatic vascular bed [J] . B. Saieag, D. Hillaire-Buys, J. Chapal, British Journal of Pharmacology . 1996,第3期

机译：腺苷5'-O-（2-硫代二磷酸）诱导大鼠胸主动脉和胰血管床松弛的机制研究
4. Guanosine stimulates neural stem cell proliferation through cAMP-CREB pathway [C] . CAIXIN SU, CAI JIANG, SHELLEY JACOBS, WSEAS international conference on medical physiology;WSEAS international conference on medical histology and embryology;WSEAS international conference on biochemistry and medical chemistry;WSEAS international conference on oncology;WSEAS international conference on medical pharmacology;WSEAS international conference on pathology;WSEAS international conference on psychiatry and psychotherapy;WSEAS international conference on cardiology;WSEAS international conference on surgery;WSEAS international conference on neurology;WSEAS international conference on gastroenterology;WSEAS international conference on ophthalmology;WSEAS international conference on obstetrics and gynaecology;WSEAS international conference on endocrinology . 2012

机译：鸟苷通过cAMP-CREB途径刺激神经干细胞增殖
5. Prostaglandin E2-induced IL-23p19 is regulated by CREB and C/EBPβ in bone marrow derived dendritic cells [D] . Kocieda, Virginia Polonia 2013

机译：前列腺素E2诱导的IL-23p19受CREB和C /EBPβ在骨髓来源的树突状细胞中的调节
6. Involvement of K+ channel permeability changes in the L-NAME and indomethacin resistant part of adenosine-5′-O-(2-thiodiphosphate)-induced relaxation of pancreatic vascular bed [O] . D Hillaire-Buys, J Chapal, N Linck, 1998

机译：K +通道通透性变化参与腺苷5-O-（2-硫代二磷酸）引起的胰血管床松弛的L-NAME和消炎痛抗性部分
7. Guanosine 5'-O-(2-thiodiphosphate) as a competitive inhibitor of GTP in hormone or cholera toxin-stimulated pancreatic adenylate cyclase [O] . Svoboda, Michal, Furnelle, Jacques, Eckstein, Fritz, 1980

机译：鸟苷5'-O-（2-硫代二磷酸）在激素或霍乱毒素刺激的胰腺腺苷酸环化酶中作为GTP的竞争性抑制剂
8. Interplay of CREB and ATF2 in Ionizing Radiation-Induced Neuroendocrine Differentiation of Prostate Cancer Cells. [R] . Hu, C., Deng, X., Hsu, C., 2012

机译：CREB和aTF2在电离辐射诱导的前列腺癌细胞神经内分泌分化中的相互作用。

Gi-protein inhibitor, guanosine 5'-O-(2-thiodiphosphate), induces senescence-associated beta-galactosidase positive cell formation through CREB phosphorylation.

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