BACKGROUND Rhabdomyosarcoma (RMS), a common soft-tissue malignancy in pediatrics, presents high invasiveness and mortality. However, besides known changes in the PAX3/7-FOXO1 fusion gene in alveolar RMS, the molecular mechanisms of the disease remain incompletely understood. The purpose of the study is to recognize potential biomarkers related with RMS and analyse their molecular mechanism, diagnosis and prognostic significance. METHODS The Gene Expression Omnibus was used to search the RMS and normal striated muscle data sets. Differentially expressed genes (DEGs) were filtered using R software. The DAVID has become accustomed to performing functional annotations and pathway analysis on DEGs. The protein interaction was constructed and further processed by the STRING tool and Cytoscape software. Kaplan-Meier was used to estimate the effect of hub genes on the ending of sarcoma sufferers, and the expression of these genes in RMS was proved by real-time polymerase chain reaction (RT-PCR). Finally, the expression of CDK1 and CCNB1 in RMS was validated by immunohistochemistry (IHC). RESULTS A total of 1932 DEGs were obtained, amongst which 1505 were up-regulated and 427were down-regulated. Up-regulated genes were largely enriched in the cell cycle, ECM-receptor interaction, PI3K/Akt and p53 pathways, whilst down-regulated genes were primarily enriched in the muscle contraction process. CDK1, CCNB1, CDC20, CCNB2, AURKB, MAD2L1, HIST2H2BE, CENPE, KIF2C and PCNA were identified as hub genes by Cytoscape analyses. Survival analysis showed that, except for HIST2H2BE, the other hub genes were highly expressed and related to poor prognosis in sarcoma. RT-PCR validation showed that CDK1, CCNB1, CDC20, CENPE and HIST2H2BE were significantly differential expression in RMS compared to the normal control. IHC revealed that the expression of CDK1 (28/32, 87.5%) and CCNB1 (26/32, 81.25%) were notably higher in RMS than normal controls (1/9, 11.1%; 0/9, 0%). Moreover, the CCNB1 was associated with the age and location of the patient's onset. CONCLUSIONS These results show that these hub genes, especially CDK1 and CCNB1, may be potential diagnostic biomarkers for RMS and provide a new perspective for the pathogenesis of RMS.

中文翻译：

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CDK1和CCNB1作为横纹肌肉瘤的潜在诊断标记：生物信息学分析后的验证。

背景技术横纹肌肉瘤（RMS）是儿科中常见的软组织恶性肿瘤，具有很高的侵袭性和死亡率。但是，除了肺泡RMS中PAX3 / 7-FOXO1融合基因的已知变化外，该疾病的分子机制仍不完全清楚。该研究的目的是识别与RMS相关的潜在生物标记，并分析其分子机制，诊断和预后意义。方法使用基因表达综合总线搜索RMS和正常横纹肌数据集。使用R软件过滤差异表达基因（DEG）。DAVID已经习惯于在DEG上执行功能注释和路径分析。通过STRING工具和Cytoscape软件构建并进一步处理蛋白质相互作用。用Kaplan-Meier来评估轮毂基因对肉瘤患者结局的影响，并通过实时聚合酶链反应（RT-PCR）证明了这些基因在RMS中的表达。最后，通过免疫组织化学（IHC）验证了RMS中CDK1和CCNB1的表达。结果共获得1932个DEG，其中上调1505个，下调427个。上调的基因在细胞周期，ECM-受体相互作用，PI3K / Akt和p53途径中大量富集，而下调的基因主要在肌肉收缩过程中富集。通过Cytoscape分析将CDK1，CCNB1，CDC20，CCNB2，AURKB，MAD2L1，HIST2H2BE，CENPE，KIF2C和PCNA鉴定为中心基因。生存分析表明，除了HIST2H2BE，其他毂基因高表达，与肉瘤的预后不良有关。RT-PCR验证显示，与正常对照相比，CDK1，CCNB1，CDC20，CENPE和HIST2H2BE在RMS中显着差异表达。IHC显示，RMS中CDK1（28/32，87.5％）和CCNB1（26/32，81.25％）的表达明显高于正常对照（1/9，11.1％; 0/9，0％）。此外，CCNB1与患者发病的年龄和位置有关。结论这些结果表明，这些中枢基因，尤其是CDK1和CCNB1，可能是RMS的潜在诊断生物标志物，并为RMS的发病机理提供了新的视角。IHC显示，RMS中CDK1（28/32，87.5％）和CCNB1（26/32，81.25％）的表达明显高于正常对照（1/9，11.1％; 0/9，0％）。此外，CCNB1与患者发病的年龄和位置有关。结论这些结果表明，这些中枢基因，尤其是CDK1和CCNB1，可能是RMS的潜在诊断生物标志物，并为RMS的发病机理提供了新的视角。IHC显示，RMS中CDK1（28/32，87.5％）和CCNB1（26/32，81.25％）的表达明显高于正常对照（1/9，11.1％; 0/9，0％）。此外，CCNB1与患者发病的年龄和位置有关。结论这些结果表明，这些中枢基因，尤其是CDK1和CCNB1，可能是RMS的潜在诊断生物标志物，并为RMS的发病机理提供了新的视角。