Zhang Tao, Chen Sixia, Peng Yi, Wang Changgang, Cheng Xi, Zhao Ren, Liu Kun
Department of General Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Front Cell Dev Biol. 2021 Oct 8;9:673873. doi: 10.3389/fcell.2021.673873. eCollection 2021.
Gene expression and alternative splicing (AS) can promote cancer development complex mechanisms. We aimed to identify and verify the hub AS events and splicing factors associated with the progression of colorectal cancer (CRC). RNA-Seq data, clinical data, and AS events of 590 CRC samples were obtained from the TCGA and TCGASpliceSeq databases. Cox univariable and multivariable analyses, KEGG, and GO pathway analyses were performed to identify hub AS events and splicing factor/spliceosome genes, which were further validated in five CRCs. In this study, we first compared differentially expressed genes and gene AS events between normal and tumor tissues. Differentially expressed genes were different from genes with differentially expressed AS events. Prognostic analysis and co-expression network analysis of gene expression and gene AS events were conducted to screen five hub gene AS events involved in CRC progression: EPB41L2, CELF2, TMEM130, VCL, and SORBS2. Using qRT-PCR, we also verified that the gene AS events SORBS2 were downregulated in tumor tissue, and gene AS events EPB41L2, CELF2, TMEM130, and VCL were upregulated in tumor tissue. The genes whose mRNA levels were significantly related to the five hub gene AS events were significantly enriched in the GO term of cell division and Notch signaling pathway. Further coexpression of gene AS events and alternative splicing factor genes revealed NOVA1 as a crucial factor regulating the hub gene AS event expression in CRC. Through experiments, we found that NOVA1 inhibited gene AS event SORBS2, which induced the migration of CRC cells the Notch pathway. Integrated analysis of gene expression and gene AS events and further experiments revealed that NOVA1-mediated SORBS2 promoted the migration of CRC, indicating its potential as a therapeutic target.
基因表达和可变剪接（AS）可通过复杂机制促进癌症发展。我们旨在识别和验证与结直肠癌（CRC）进展相关的关键AS事件和剪接因子。从TCGA和TCGASpliceSeq数据库中获取了590个CRC样本的RNA-Seq数据、临床数据和AS事件。进行Cox单变量和多变量分析、KEGG和GO通路分析以识别关键AS事件和剪接因子/剪接体基因，并在五个CRC样本中进一步验证。在本研究中，我们首先比较了正常组织和肿瘤组织之间的差异表达基因和基因AS事件。差异表达基因与具有差异表达AS事件的基因不同。对基因表达和基因AS事件进行预后分析和共表达网络分析，以筛选出参与CRC进展的五个关键基因AS事件：EPB41L2、CELF2、TMEM130、VCL和SORBS2。使用qRT-PCR，我们还验证了基因AS事件SORBS2在肿瘤组织中下调，而基因AS事件EPB41L2、CELF2、TMEM130和VCL在肿瘤组织中上调。其mRNA水平与这五个关键基因AS事件显著相关的基因在细胞分裂和Notch信号通路的GO术语中显著富集。基因AS事件和可变剪接因子基因的进一步共表达揭示了NOVA1是调节CRC中关键基因AS事件表达的关键因子。通过实验，我们发现NOVA1抑制基因AS事件SORBS2，这通过Notch途径诱导CRC细胞迁移。基因表达和基因AS事件的综合分析以及进一步实验表明，NOVA1介导的SORBS2促进了CRC的迁移，表明其作为治疗靶点的潜力。
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