Department of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, China.
Department of Radiology, Affiliated Hospital of Yunnan University, Kunming, China.
PeerJ. 2023 Nov 7;11:e16312. doi: 10.7717/peerj.16312. eCollection 2023.
Alternative splicing (AS) is a biological process that allows genes to be translated into diverse proteins. However, aberrant AS can predispose cells to aberrations in biological mechanisms. RNA binding proteins (RBPs), closely affiliated with AS, have gained increased attention in recent years. Among these RBPs, RBM25 has been reported to participate in the cardiac pathological mechanism through regulating AS; however, the involvement of RBM25 as a splicing factor in heart failure remains unclarified.
RBM25 was overexpressed in H9c2 cells to explore the target genes bound and regulated by RBM25 during heart failure. RNA sequencing (RNA-seq) was used to scrutinize the comprehensive transcriptional level before identifying AS events influenced by RBM25. Further, improved RNA immunoprecipitation sequencing (iRIP-seq) was employed to pinpoint RBM25-binding sites, and RT-qPCR was used to validate specific genes modulated by RBM25.
RBM25 was found to upregulate the expression of genes pertinent to the inflammatory response and viral processes, as well as to mediate the AS of genes associated with cellular apoptosis and inflammation. Overlap analysis between RNA-seq and iRIP-seq suggested that RBM25 bound to and manipulated the AS of genes associated with inflammation in H9c2 cells. Moreover, qRT-PCR confirmed , , and as the binding and AS regulatory targets of RBM25.
Our research implies that RBM25 plays a contributory role in cardiac inflammatory responses via its ability to bind to and regulate the AS of related genes. This study offers preliminary evidence of the influence of RBM25 on inflammation in H9c2 cells.
可变剪接（AS）是一种使基因能够翻译成不同蛋白质的生物学过程。然而，异常的 AS 会使细胞容易发生生物学机制的异常。近年来，与 AS 密切相关的 RNA 结合蛋白（RBPs）受到了越来越多的关注。在这些 RBPs 中，已经有报道称 RBM25 通过调节 AS 参与心脏病理机制；然而，RBM25 作为心力衰竭中的剪接因子的参与仍不清楚。
在 H9c2 细胞中过表达 RBM25，以探讨心力衰竭期间 RBM25 结合和调节的靶基因。使用 RNA 测序（RNA-seq）来仔细检查 RBM25 影响之前的全面转录水平。进一步，采用改进的 RNA 免疫沉淀测序（iRIP-seq）来确定 RBM25 结合位点，并采用 RT-qPCR 来验证 RBM25 调节的特定基因。
发现 RBM25 上调与炎症反应和病毒过程相关的基因表达，并介导与细胞凋亡和炎症相关的基因的 AS。RNA-seq 和 iRIP-seq 的重叠分析表明，RBM25 结合并调节了 H9c2 细胞中与炎症相关的基因的 AS。此外，qRT-PCR 证实 、 和 是 RBM25 的结合和 AS 调节靶标。
我们的研究表明，RBM25 通过结合和调节相关基因的 AS，在心脏炎症反应中发挥作用。本研究为 RBM25 对 H9c2 细胞炎症的影响提供了初步证据。
