Mevalonate cascade regulation of airway mesenchymal cell autophagy and apoptosis: a dual role for p53.
Mevalonate cascade regulation of airway mesenchymal cell autophagy and apoptosis: a dual role for p53.
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DOI：
10.1371/journal.pone.0016523
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发表时间：
2011-01-31
期刊：
影响因子：
3.7
通讯作者：
Halayko AJ
中科院分区：
文献类型：
--
作者：
Ghavami S;Mutawe MM;Sharma P;Yeganeh B;McNeill KD;Klonisch T;Unruh H;Kashani HH;Schaafsma D;Los M;Halayko AJ
他汀类药物抑制胆固醇生物合成的近端步骤，并与各种疾病（包括癌症和肺部疾病）的健康益处有关。我们以前研究了他汀类药物在气道间充质细胞中引发的凋亡途径，并确定减少小GTP酶的异戊烯化是导致p53介导的细胞死亡的主要效应机制。在这里，我们通过评估细胞凋亡和自噬的终点，并研究它们的相互作用和同步调节，扩展了我们对他汀类药物诱导的细胞死亡的研究。使用原代培养的人气道平滑肌（HASM）和人气道成纤维细胞（HAF），通过透射电子显微镜、细胞化学（溶酶体数量和与LC 3的共定位）和免疫印迹（LC 3脂质化和Atg 12 -5复合物形成）评估自噬和自噬体形成和流量。自噬的化学抑制增加辛伐他汀诱导的半胱天冬酶活化和细胞死亡。类似地，用shRNA沉默Atg 5，从而防止Atg 5 -12复合物形成，增加了辛伐他汀的促凋亡作用。辛伐他汀同时增加p53依赖性表达的p53上调的凋亡调节剂（p53 up-regulated modulator of apoptosis，PDMA），NOXA和损伤调节的自噬调节剂（damage-regulated autophagy modulator，DRAM）。值得注意的是，甲羟戊酸级联抑制诱导的自噬和细胞凋亡均为p53依赖性：辛伐他汀增加细胞核p53蓄积，环匹林-α和p53 shRNAi均部分抑制NOXA、caspase-3/7裂解（细胞凋亡）和DRAM表达、Atg 5 -12复合物形成、LC 3脂化和自噬体形成（自噬）。此外，自噬反应被迅速诱导，显著延迟细胞凋亡，表明存在时间协调的p53调控网络。这些发现与他汀类药物治疗阻塞性气道疾病的方法的发展有关。
Statins inhibit the proximal steps of cholesterol biosynthesis, and are linked to health benefits in various conditions, including cancer and lung disease. We have previously investigated apoptotic pathways triggered by statins in airway mesenchymal cells, and identified reduced prenylation of small GTPases as a primary effector mechanism leading to p53-mediated cell death. Here, we extend our studies of statin-induced cell death by assessing endpoints of both apoptosis and autophagy, and investigating their interplay and coincident regulation. Using primary cultured human airway smooth muscle (HASM) and human airway fibroblasts (HAF), autophagy, and autophagosome formation and flux were assessed by transmission electron microscopy, cytochemistry (lysosome number and co-localization with LC3) and immunoblotting (LC3 lipidation and Atg12-5 complex formation). Chemical inhibition of autophagy increased simvastatin-induced caspase activation and cell death. Similarly, Atg5 silencing with shRNA, thus preventing Atg5-12 complex formation, increased pro-apoptotic effects of simvastatin. Simvastatin concomitantly increased p53-dependent expression of p53 up-regulated modulator of apoptosis (PUMA), NOXA, and damage-regulated autophagy modulator (DRAM). Notably both mevalonate cascade inhibition-induced autophagy and apoptosis were p53 dependent: simvastatin increased nuclear p53 accumulation, and both cyclic pifithrin-α and p53 shRNAi partially inhibited NOXA, PUMA expression and caspase-3/7 cleavage (apoptosis) and DRAM expression, Atg5-12 complex formation, LC3 lipidation, and autophagosome formation (autophagy). Furthermore, the autophagy response is induced rapidly, significantly delaying apoptosis, suggesting the existence of a temporally coordinated p53 regulation network. These findings are relevant for the development of statin-based therapeutic approaches in obstructive airway disease.
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DOI：
10.1152/ajplung.00013.2006
发表时间：
2006-09-01
影响因子：
4.9
作者：
Gosens, Reinoud;Stelmack, Gerald L.;Halayko, Andrew J.
通讯作者：
Halayko, Andrew J.
影响因子：
13.3
作者：
Crighton, Diane;Wilkinson, Simon;Ryan, Kevin M.
通讯作者：
Ryan, Kevin M.
影响因子：
5.3
作者：
Ghavami S;Asoodeh A;Klonisch T;Halayko AJ;Kadkhoda K;Kroczak TJ;Gibson SB;Booy EP;Naderi-Manesh H;Los M
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Los M
影响因子：
8
作者：
Iwamaru, A.;Kondo, Y.;Kondo, S.
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Kondo, S.
影响因子：
5.3
作者：
Kaneta, S;Satoh, K;Ichihara, K
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Ichihara, K