Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Division of Hematopoiesis, Joint Research Center for Human Retrovirus Infection and Graduate School Medical Sciences, Kumamoto University, Kumamoto 860-0811, Japan.
Int J Mol Sci. 2024 Sep 22;25(18):10170. doi: 10.3390/ijms251810170.
We have previously shown that the overexpression of acetyl-CoA carboxylase 1 (ACC1) was associated with the poor prognosis of cholangiocarcinoma (CCA) patients, and suppression of its expression in CCA cell lines deteriorated cell growth. The present study explored the mechanism by which ACC1 inhibition affects global protein acetylation, using genetic knockdown and pharmacological inhibition with an ACC1 inhibitor ND-646 as models. Both ACC1 knockdown and ACC1-inhibitor-treated cells displayed the hyperacetylation of proteins, accompanied by impaired growth and migration. The immunoprecipitation of hyperacetylated proteins using the anti-acetylated lysine antibody, followed by tandem mass spectrometry, identified three potential verification candidates, namely POTE ankyrin domain family member E, peroxisomal biogenesis factor 1, and heat shock protein 90 beta (HSP90B). HSP90 acetylation was the candidate selected for the verification of protein acetylation. To establish the effects of protein hyperacetylation, treatment with suberoylanilide hydroxamic acid (SAHA), a lysine deacetylase inhibitor, was conducted, and this served as an independent model. Decreased tumor growth but increased acetylated protein levels were observed in ACC1-KD xenograft tumors. Hyperacetylated-alleviated cell growth and migration were consistently observed in the SAHA-treated models. The molecular linkage between protein hyperacetylation and the AKT/GSK3β/Snail pathway was demonstrated. This study highlighted the importance of protein acetylation in CCA progression, suggesting that ACC1 and KDAC are potential targets for CCA treatment.
我们之前已经表明，乙酰辅酶 A 羧化酶 1（ACC1）的过表达与胆管癌（CCA）患者的预后不良有关，并且在 CCA 细胞系中抑制其表达会恶化细胞生长。本研究通过基因敲低和 ACC1 抑制剂 ND-646 的药理学抑制来探讨 ACC1 抑制影响全局蛋白质乙酰化的机制，作为模型。ACC1 敲低和 ACC1 抑制剂处理的细胞均表现出蛋白质的过度乙酰化，伴随生长和迁移受损。使用抗乙酰化赖氨酸抗体对过度乙酰化蛋白进行免疫沉淀，然后进行串联质谱分析，鉴定出三个潜在的验证候选物，即 POTE 锚蛋白域家族成员 E、过氧化物酶体生物发生因子 1 和热休克蛋白 90β（HSP90B）。HSP90 乙酰化是选择用于验证蛋白质乙酰化的候选物。为了建立蛋白质过度乙酰化的影响，用赖氨酸去乙酰化酶抑制剂 suberoylanilide hydroxamic acid（SAHA）进行处理，作为独立模型。在 ACC1-KD 异种移植肿瘤中观察到肿瘤生长减少但乙酰化蛋白水平增加。在 SAHA 处理的模型中观察到过度乙酰化缓解细胞生长和迁移。证明了蛋白质过度乙酰化与 AKT/GSK3β/Snail 通路之间的分子联系。这项研究强调了蛋白质乙酰化在 CCA 进展中的重要性，表明 ACC1 和 KDAC 是 CCA 治疗的潜在靶点。
