The antiparasitic agent ivermectin offers more promises to treat a diverse range of diseases. However, a comprehensive proteomic analysis of ivermectin‐treated ovarian cancer (OC) cells has not been performed. This study sought to identify ivermectin‐related proteomic profiling and molecular network alterations in human OC cells. Stable isotope labeling with amino acids in cell culture (SILAC)‐based quantitative proteomics was used to study the human OC TOV‐21G cells. After TOV‐21G cells underwent 10 passages in SILAC‐labeled growth media, TOV‐21G cells were treated with 10 ml of 20 μmol/L ivermectin in cell growing medium for 24 h. The SILAC‐labeled proteins were digested with trypsin; tryptic peptides were identified with mass spectrometry (MS). Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was used to mine signaling pathway alterations with ivermectin‐related proteins in TOV‐21G cells. Gene ontology (GO) analysis was used to explore biological functions of ivermectin‐related proteins, including biological processes (BPs), cellular components (CCs), and molecular functions (MFs). The protein‐protein interaction network was analyzed with molecular complex detection (MCODE) to identify hub modules. In total, 4,447 proteins were identified in ivermectin‐treated TOV‐21G cells. KEGG analysis revealed 89 statistically significant signaling pathways. Interestingly, the clustering analysis of these pathways showed that ivermectin was involved in various cancer pathogenesis processes, including modulation of replication, RNA metabolism, and translational machinery. GO analysis revealed 69 statistically significant CCs, 87 MFs, and 62 BPs. Furthermore, MCODE analysis identified five hub modules, including 147 hub molecules. Those hub modules involved ribosomal proteins, RNA‐binding proteins, cell‐cycle progression‐related proteins, proteasome subunits, and minichromosome maintenance proteins. These findings demonstrate that SILAC quantitative proteomics is an effective method to analyze ivermectin‐treated cells, provide the first ivermectin‐related proteomic profiling and molecular network alterations in human OC cells, and provide deeper insights into molecular mechanisms and functions of ivermectin to inhibit OC cells.

中文翻译：

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人卵巢癌细胞中伊维菌素相关蛋白质组学分析和分子网络改变的 SILAC 定量蛋白质组学分析

抗寄生虫药伊维菌素为治疗多种疾病提供了更多希望。然而，尚未对伊维菌素处理的卵巢癌 (OC) 细胞进行全面的蛋白质组学分析。本研究旨在确定人类 OC 细胞中伊维菌素相关的蛋白质组学分析和分子网络改变。基于细胞培养中氨基酸的稳定同位素标记 (SILAC) 定量蛋白质组学被用于研究人类 OC TOV-21G 细胞。TOV-21G 细胞在 SILAC 标记的生长培养基中传代 10 次后，TOV-21G 细胞在细胞生长培养基中用 10 ml 20 μmol/L 伊维菌素处理 24 小时。SILAC 标记的蛋白质用胰蛋白酶消化；用质谱法 (MS) 鉴定胰蛋白酶肽。京都基因和基因组百科全书 (KEGG) 分析用于挖掘 TOV-21G 细胞中伊维菌素相关蛋白的信号通路改变。基因本体（GO）分析用于探索伊维菌素相关蛋白的生物学功能，包括生物学过程（BPs）、细胞成分（CCs）和分子功能（MFs）。使用分子复合物检测（MCODE）分析蛋白质-蛋白质相互作用网络以识别枢纽模块。总共在伊维菌素处理的 TOV-21G 细胞中鉴定了 4,447 种蛋白质。KEGG 分析揭示了 89 条具有统计学意义的信号通路。有趣的是，对这些通路的聚类分析表明，伊维菌素参与了多种癌症发病机制，包括复制调节、RNA 代谢和翻译机制。GO 分析显示 69 个具有统计学意义的 CC、87 个 MF 和 62 个 BP。此外，MCODE 分析确定了五个中枢模块，包括 147 个中枢分子。这些枢纽模块涉及核糖体蛋白、RNA 结合蛋白、细胞周期进程相关蛋白、蛋白酶体亚基和微染色体维持蛋白。这些发现表明，SILAC 定量蛋白质组学是分析伊维菌素处理细胞的有效方法，提供了第一个伊维菌素相关蛋白质组学分析和人类 OC 细胞的分子网络改变，并提供了对伊维菌素抑制 OC 细胞的分子机制和功能的更深入的了解. RNA结合蛋白、细胞周期进程相关蛋白、蛋白酶体亚基和微染色体维持蛋白。这些发现表明 SILAC 定量蛋白质组学是分析伊维菌素处理细胞的有效方法，提供了第一个伊维菌素相关蛋白质组学分析和人类 OC 细胞的分子网络改变，并提供了对伊维菌素抑制 OC 细胞的分子机制和功能的更深入的了解. RNA 结合蛋白、细胞周期进程相关蛋白、蛋白酶体亚基和微染色体维持蛋白。这些发现表明，SILAC 定量蛋白质组学是分析伊维菌素处理细胞的有效方法，提供了第一个伊维菌素相关蛋白质组学分析和人类 OC 细胞的分子网络改变，并提供了对伊维菌素抑制 OC 细胞的分子机制和功能的更深入的了解.