未经授权:睾丸生殖细胞肿瘤(TGCT)是年轻男性中最常见的肿瘤,但是分子特征,特别是其亚型之间的选择性剪接(AS)尚未被探索。
未经评估:为了研究TGCT亚型之间的差异,我们全面分析了基因表达的数据,交替拼接(AS),和来自TCGA数据库的TGCT患者的体细胞突变。利用基因本体论(GO)富集分析分别探讨差异表达基因和剪接基因的功能,并进行Spearman相关分析,探讨差异基因与AS事件的相关性。此外,集合数据库转录图集详细阐述了AS调节基因表达的可能模式。And,我们鉴定了调节基因表达和AS的重要转录因子,并在TGCT细胞系中对其进行了功能验证.
UNASSIGNED:我们发现在胚胎癌和精原细胞瘤中表达与AS之间存在显著差异,而混合细胞肿瘤介于两者之间。GO富集分析显示,差异表达和剪接基因都在转录调控途径中富集,并确定表达与AS事件之间存在明显的相关性。通过分析转录图和剪接发生的位点,我们已经证明AS以多种方式调节基因表达。我们进一步确定了两个参与AS调节的枢轴AS相关分子(SOX2和HDAC9),在胚胎癌和精原细胞瘤细胞系中得到验证。亚型之间的体细胞突变差异也令人担忧,我们的结果表明,某些基因(B3GNT8,CAPN7,FAT4,GRK1,TACC2和TRAM1L1)的突变仅发生在胚胎癌中,而KIT中的突变,KARS,NRAS仅在精原细胞瘤中观察到。
未经批准:总而言之,我们的分析揭示了基因表达的差异,AS和TGCT亚型的体细胞突变,为TGCT患者的临床诊断和精准治疗提供分子依据。
Testicular germ cell tumor (TGCT) is the most common tumor in young men, but molecular signatures, especially the alternative splicing (AS) between its subtypes have not yet been explored.
To investigate the differences between TGCT subtypes, we comprehensively analyzed the data of gene expression, alternative splicing (AS), and somatic mutation in TGCT patients from the TCGA database. The gene ontology (GO) enrichment analyses were used to explore the function of differentially expressed genes and spliced genes respectively, and Spearman correlation analysis was performed to explore the correlation between differential genes and AS events. In addition, the possible patterns in which AS regulates gene expression were elaborated by the ensemble database transcript atlas. And, we identified important transcription factors that regulate gene expression and AS and functionally validated them in TGCT cell lines.
We found significant differences between expression and AS in embryonal carcinoma and seminoma, while mixed cell tumors were in between. GO enrichment analyses revealed that both differentially expressed and spliced genes were enriched in transcriptional regulatory pathways, and obvious correlation between expression and AS events was determined. By analyzing the transcript map and the sites where splicing occurs, we have demonstrated that AS regulates gene expression in a variety of ways. We further identified two pivot AS-related molecules (SOX2 and HDAC9) involved in AS regulation, which were validated in embryonal carcinoma and seminoma cell lines. Differences in somatic mutations between subtypes are also of concern, with our results suggesting that mutations in some genes (B3GNT8, CAPN7, FAT4, GRK1, TACC2, and TRAM1L1) occur only in embryonal carcinoma, while mutations in KIT, KARS, and NRAS are observed only in seminoma.
In conclusion, our analysis revealed the differences in gene expression, AS and somatic mutation among TGCT subtypes, providing a molecular basis for clinical diagnosis and precise therapy of TGCT patients.