在之前使用 iTRAQ（相对和绝对定量同量异位标签）技术的研究中，我们在乳腺癌组织中鉴定出了一系列表现出高水平乙酰化的蛋白质。尽管取得了这一进展，但这些乙酰化蛋白在癌症生物学背景下的具体功能和含义尚未阐明。本研究旨在系统地研究这些乙酰化蛋白的功能作用，旨在确定乳腺癌病理生理学中潜在的治疗靶点。通过生物信息学鉴定乙酰化靶点，并随后确认其表达和乙酰化。蛋白质组分析和验证研究确定了潜在的乙酰转移酶和脱乙酰酶。我们通过催化活性、葡萄糖消耗、ATP 水平和乳酸生成测定来评估代谢功能。通过活力、循环分析、克隆形成测定、PCNA 摄取、伤口愈合、Transwell 测定和 MMP/EMT 标记物检测来评估细胞增殖和转移。乳腺癌中的乙酰化蛋白主要参与代谢，显着影响糖酵解和三羧酸循环。值得注意的是，PGK1 在赖氨酸 323 处表现出最高的乙酰化，并且在乳腺癌组织中表现出表达和乙酰化增加，特别是在 T47D 和 MCF-7 细胞中。值得注意的是，在 T47D 细胞中鉴定出 18 种乙酰转移酶或脱乙酰酶，其中 p300 和 Sirtuin3 已被验证与 PGK1 的相互作用。 323 K 的乙酰化通过提高其活性、葡萄糖摄取、ATP 产生和乳酸输出来增强 PGK1 的代谢作用。这种修饰还促进了细胞增殖，活力、S 期比例、克隆性和 PCNA 水平的增加证明了这一点。此外，PGK1-323 K 乙酰化促进转移，改善伤口愈合、细胞侵袭，上调 MMP2、MMP9、N-钙粘蛋白和波形蛋白，同时下调 E-钙粘蛋白。PGK1-323 K 乙酰化在 T47D 和 MCF-7 管腔中显着升高在乳腺癌细胞中，这种乙酰化可以通过 p300 和 Sirtuin3 进行调节。 PGK1-323 K 乙酰化促进细胞糖酵解、增殖和转移，突出了乳腺癌治疗的新表观遗传靶标。© 2024。作者。

In prior research employing iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) technology, we identified a range of proteins in breast cancer tissues exhibiting high levels of acetylation. Despite this advancement, the specific functions and implications of these acetylated proteins in the context of cancer biology have yet to be elucidated. This study aims to systematically investigate the functional roles of these acetylated proteins with the objective of identifying potential therapeutic targets within breast cancer pathophysiology.Acetylated targets were identified through bioinformatics, with their expression and acetylation subsequently confirmed. Proteomic analysis and validation studies identified potential acetyltransferases and deacetylases. We evaluated metabolic functions via assays for catalytic activity, glucose consumption, ATP levels, and lactate production. Cell proliferation and metastasis were assessed through viability, cycle analysis, clonogenic assays, PCNA uptake, wound healing, Transwell assays, and MMP/EMT marker detection.Acetylated proteins in breast cancer were primarily involved in metabolism, significantly impacting glycolysis and the tricarboxylic acid cycle. Notably, PGK1 showed the highest acetylation at lysine 323 and exhibited increased expression and acetylation across breast cancer tissues, particularly in T47D and MCF-7 cells. Notably, 18 varieties acetyltransferases or deacetylases were identified in T47D cells, among which p300 and Sirtuin3 were validated for their interaction with PGK1. Acetylation at 323 K enhanced PGK1's metabolic role by boosting its activity, glucose uptake, ATP production, and lactate output. This modification also promoted cell proliferation, as evidenced by increased viability, S phase ratio, clonality, and PCNA levels. Furthermore, PGK1-323 K acetylation facilitated metastasis, improving wound healing, cell invasion, and upregulating MMP2, MMP9, N-cadherin, and Vimentin while downregulating E-cadherin.PGK1-323 K acetylation was significantly elevated in T47D and MCF-7 luminal A breast cancer cells and this acetylation could be regulated by p300 and Sirtuin3. PGK1-323 K acetylation promoted cell glycolysis, proliferation, and metastasis, highlighting novel epigenetic targets for breast cancer therapy.© 2024. The Author(s).