Kim Jisu, Lee Kang Pa, Kim Bom Sahn, Lee Sang Ju, Moon Byung Seok, Baek Suji
Department of Sports Medicine and Science in Graduate School, Konkuk University, Seoul 05029, Korea.
Research & Development Center, UMUST R&D Corporation, Seoul 05029, Korea.
Korean J Physiol Pharmacol. 2020 May 1;24(3):241-248. doi: 10.4196/kjpp.2020.24.3.241.
Luminespib (AUY922), a heat shock proteins 90 inhibitor, has anti-neoplastic and antitumor effects. However, it is not clear whether AUY922 affects events in vascular diseases. We investigated the effects of AUY922 on the platelet-derived growth factor (PDGF)-BB-stimulated proliferation and migration of vascular smooth muscle cells (VSMC). VSMC viability was detected using the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)--tetrazolium-5-carboxanilide) reagent. To detect the attenuating effects of AUY922 on PDGF-BB-induced VSMCs migration , we performed the Boyden chamber and scratch wound healing assays. To identify AUY922-mediated changes in the signaling pathway, the phosphorylation of protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) 1/2 was analyzed by immunoblotting. The inhibitory effects of AUY922 on migration and proliferation were tested using an aortic ring assay. AUY922 was not cytotoxic at concentrations up to 5 nM. PDGF-BB-induced VSMC proliferation, migration, and sprout outgrowth were significantly decreased by AUY922 in a dose-dependent manner. AUY922 significantly reduced the PDGF-BB-stimulated phosphorylation of Akt and ERK1/2. Furthermore, PD98059 (a selective ERK1/2 inhibitor) and LY294002 (a selective Akt inhibitor) decreased VSMC migration and proliferation by inhibiting phosphorylation of Akt and ERK1/2. Greater attenuation of PDGF-BB-induced cell viability and migration was observed upon treatment with PD98059 or LY294002 in combination with AUY922. AUY922 showed anti-proliferation and anti-migration effects towards PDGF-BBinduced VSMCs by regulating the phosphorylation of ERK1/2 and Akt. Thus, AUY922 is a candidate for the treatment of atherosclerosis and restenosis.
17-烯丙胺基-17-去甲氧基格尔德霉素（AUY922）是一种热休克蛋白90抑制剂，具有抗肿瘤和抗癌作用。然而，AUY922是否影响血管疾病相关事件尚不清楚。我们研究了AUY922对血小板衍生生长因子（PDGF）-BB刺激的血管平滑肌细胞（VSMC）增殖和迁移的影响。使用XTT（2,3-双-(2-甲氧基-4-硝基-5-磺基苯基)-5-羧基苯胺）试剂检测VSMC活力。为了检测AUY922对PDGF-BB诱导的VSMC迁移的减弱作用，我们进行了博伊登小室实验和划痕伤口愈合实验。为了确定AUY922介导的信号通路变化，通过免疫印迹分析蛋白激酶B（Akt）和细胞外信号调节激酶（ERK）1/2的磷酸化情况。使用主动脉环实验测试AUY922对迁移和增殖的抑制作用。在浓度高达5 nM时，AUY922没有细胞毒性。AUY922以剂量依赖性方式显著降低PDGF-BB诱导的VSMC增殖、迁移和芽生。AUY922显著降低了PDGF-BB刺激的Akt和ERK1/2磷酸化。此外，PD98059（一种选择性ERK1/2抑制剂）和LY294002（一种选择性Akt抑制剂）通过抑制Akt和ERK1/2的磷酸化降低了VSMC迁移和增殖。在用PD98059或LY294002与AUY922联合处理后，观察到对PDGF-BB诱导的细胞活力和迁移有更大程度的减弱。AUY922通过调节ERK1/2和Akt的磷酸化，对PDGF-BB诱导的VSMC表现出抗增殖和抗迁移作用。因此，AUY922是治疗动脉粥样硬化和再狭窄的候选药物。
