Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Borwell Building 644E, Lebanon, New Hampshire 03756, United States.
Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania Medical School, 421 Curie Blvd. Room 612 BRB II/III, Philadelphia, Pennsylvania 19104, United States.
J Proteome Res. 2023 Sep 1;22(9):3040-3053. doi: 10.1021/acs.jproteome.3c00340. Epub 2023 Aug 8.
Tamoxifen (TMX), a selective estrogen receptor modulator, is commonly used in the treatment of hormone-responsive cancers. However, the effects of TMX in anabolic tissues harboring estrogen receptors, such as skeletal muscle, are poorly understood. We report a tandem mass-tag approach to TMX-treated myogenesis in CC cells, a well-characterized model of murine skeletal muscle differentiation. A longitudinal analysis of >10,000 proteins identified in untreated CC myogenesis revealed a novel subset of 1,062 myogenically regulated proteins. These proteins clustered into five distinct longitudinal expression trends which significantly overlap those obtained in similar analyses performed in human myocytes. We document a specific functional enrichment for adiponectin-signaling unique to TMX-treated myogenesis, as well as a subset of 198 proteins that are differentially expressed in TMX-treated cells relative to controls at one or more stages of myogenesis, the majority of which were involved in steroid and lipid metabolism. Further analysis highlights metallothionein-1 as a novel target of TMX treatment at each stage of CC myogenesis. Finally, we present a powerful, self-validating pipeline for analyzing the total proteomic response to treatment across every stage of muscle cell development which can be easily adapted to study the effects of other drugs on myogenesis.
他莫昔芬（TMX）是一种选择性雌激素受体调节剂，常用于治疗激素反应性癌症。然而，TMX 在含有雌激素受体的合成组织（如骨骼肌）中的作用知之甚少。我们报告了一种串联质量标签方法，用于研究 CC 细胞中的 TMX 诱导的成肌作用，CC 细胞是一种成熟的鼠骨骼肌分化模型。对未经处理的 CC 成肌作用中鉴定的>10000 种蛋白质进行的纵向分析显示，存在一组新的 1062 种肌生成调节蛋白。这些蛋白质聚类为五个不同的纵向表达趋势，与在人类肌细胞中进行的类似分析获得的趋势显著重叠。我们记录了一个独特的脂联素信号的特定功能富集，该信号仅存在于 TMX 处理的成肌作用中，以及一组在 TMX 处理的细胞中与对照组在成肌作用的一个或多个阶段差异表达的 198 种蛋白质，其中大多数涉及类固醇和脂质代谢。进一步的分析强调了金属硫蛋白-1 作为 TMX 治疗在 CC 成肌作用的每个阶段的新靶点。最后，我们提出了一种强大的、自我验证的管道，用于分析在肌肉细胞发育的每个阶段对治疗的总蛋白质组反应，该管道可以很容易地适应研究其他药物对成肌作用的影响。
