Transcriptomic analysis of Cr(VI)-induced changes in C2C12 cells during myogenic differentiation

IF 3.6 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of Trace Elements in Medicine and Biology Pub Date : 2025-08-09 DOI:10.1016/j.jtemb.2025.127714

Sun Young Park, Hong Sun

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引用次数: 0

Abstract

Background

Hexavalent chromium [Cr(VI)] is an environmental toxicant extensively used in a variety of industrial processes including chrome plating, leather tanning, textile manufacturing, aircraft production, and stainless-steel production. Our previous study reported that exposure to Cr(VI) inhibited C2C12 myogenic differentiation in a dose-dependent manner, yet the transcriptional mechanisms underlying Cr(VI)-induced disruption of myogenesis remains poorly understood. This study aimed to characterize the global transcriptional alterations during C2C12 myogenic differentiation and identify molecular pathways disrupted upon Cr(VI) exposure.

Methods

C2C12 cells were differentiated in the presence of 0, 2, or 5 μM of Cr(VI) and collected at differentiation days 0, 1, 2, and 4. Whole transcriptome analysis of a total of 30 samples (with 3 biological replicates per condition) was performed using RNA-sequencing followed by differential gene expression analysis, unsupervised fuzzy c-means clustering, GO biological processes functional annotation, and KEGG pathway enrichment analysis.

Results

Cr(VI) exposure resulted in a massive transcriptomic change in differentiating C2C12 cells. Fuzzy c-means clustering identified 12 distinct gene expression patterns, with Clusters 3, 10, and 12 showing significant overlap with Cr(VI)-regulated genes. Functional enrichment analyses revealed Cr(VI) alters genes involved in early-stage cell cycle regulation and DNA repair as well as terminal differentiation processes like sarcomere organization and muscle contraction. Specifically, Cr(VI) suppressed expression of key structural and contractile genes and disrupted pathways essential for myogenic differentiation, cell cycle regulation and DNA damage repair. Furthermore, Cr(VI) disrupted Hippo signaling by downregulating Tead4 and its downstream myogenic targets such as MyoG, Cav3, Mustn1, and Dysf suggesting a mechanism for impaired differentiation.

Conclusion

This study highlights the widespread alterations of Cr(VI) exposure on transcriptional programs – including structural development, genomic stability, and cell cycle withdrawal – governing muscle development and maturation, offering insight into how Cr(VI) exposure affects skeletal muscle health.

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Cr（VI）诱导C2C12细胞在成肌分化过程中的转录组学分析

六价铬[Cr(VI)]是一种环境毒物，广泛用于各种工业过程，包括镀铬，皮革鞣制，纺织制造，飞机生产和不锈钢生产。我们之前的研究报道了暴露于Cr（VI）以剂量依赖的方式抑制C2C12肌生成分化，但Cr（VI）诱导的肌生成中断的转录机制仍然知之甚少。本研究旨在表征C2C12肌源性分化过程中的全局转录改变，并确定暴露于Cr（VI）后被破坏的分子通路。方法sc2c12细胞分别在0、2、5 μM的Cr（VI）作用下进行分化，并于分化第0、1、2、4天收集。通过rna测序、差异基因表达分析、无监督模糊c均值聚类、GO生物过程功能注释和KEGG通路富集分析，对30个样本（每种条件下3个生物重复）进行了全转录组分析。结果scr （VI）暴露导致分化C2C12细胞的大量转录组变化。模糊c均值聚类鉴定出12种不同的基因表达模式，其中集群3、10和12与Cr（VI）调控基因显著重叠。功能富集分析显示，Cr（VI）改变了参与早期细胞周期调控和DNA修复以及终端分化过程（如肌节组织和肌肉收缩）的基因。具体来说，Cr（VI）抑制了关键结构和收缩基因的表达，并破坏了肌源性分化、细胞周期调节和DNA损伤修复所必需的途径。此外，Cr（VI）通过下调Tead4及其下游肌生成靶点（如MyoG、Cav3、Mustn1和Dysf）来破坏Hippo信号通路，提示分化受损的机制。本研究强调了Cr（VI）暴露对调控肌肉发育和成熟的转录程序（包括结构发育、基因组稳定性和细胞周期退出）的广泛改变，为Cr（VI）暴露如何影响骨骼肌健康提供了见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Trace Elements in Medicine and Biology 医学-内分泌学与代谢

CiteScore

6.60

自引率

2.90%

发文量

202

审稿时长

85 days

期刊介绍： The journal provides the reader with a thorough description of theoretical and applied aspects of trace elements in medicine and biology and is devoted to the advancement of scientific knowledge about trace elements and trace element species. Trace elements play essential roles in the maintenance of physiological processes. During the last decades there has been a great deal of scientific investigation about the function and binding of trace elements. The Journal of Trace Elements in Medicine and Biology focuses on the description and dissemination of scientific results concerning the role of trace elements with respect to their mode of action in health and disease and nutritional importance. Progress in the knowledge of the biological role of trace elements depends, however, on advances in trace elements chemistry. Thus the Journal of Trace Elements in Medicine and Biology will include only those papers that base their results on proven analytical methods. Also, we only publish those articles in which the quality assurance regarding the execution of experiments and achievement of results is guaranteed.