CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis
CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis
复制标题
CDK1 和 CCNB1 作为横纹肌肉瘤的潜在诊断标志物：生物信息学分析后的验证
DOI：
10.1186/s12920-019-0645-x
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发表时间：
2019-12-23
影响因子：
2.7
通讯作者：
Li, Feng
中科院分区：
文献类型：
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作者：
Li, Qianru;Zhang, Liang;Li, Feng
横纹肌肉瘤(RMS)是儿科常见的软组织恶性肿瘤，具有很高的侵袭性和死亡率。然而，除了肺泡RMS中PAX3/7-FOXO1融合基因的已知变化外，该病的分子机制仍不完全清楚。本研究的目的是识别与RMS相关的潜在生物标志物，并分析它们的分子机制、诊断和预后意义。方法使用基因表达总表搜索RMS和正常横纹肌数据集。用R软件筛选差异表达基因(Deg)。David已经习惯于对DEG进行功能注释和路径分析，蛋白质相互作用是通过StringTool和Cytosscape软件构建和进一步处理的。Kaplan-Meier法检测HUB基因在肉瘤患者结局中的作用，实时定量聚合酶链式反应(RT-PCR)检测这些基因在RMS中的表达。结果共获得1932个DEG，其中上调1505个，下调427个。上调的基因主要集中在细胞周期、ECM-受体相互作用、PI3K/Akt和P53途径中，而下调的基因主要集中在肌肉收缩过程中。细胞角蛋白分析确定CDK1、CCNB1、CDC20、CCNB2、AURKB、MAD2L1、HIST2H2BE、CENPE、KIF2C和PCNA为HUB基因。生存分析显示，除HIST2H2BE外，其余HUB基因均高表达，且与肉瘤预后不良有关。RT-PCR验证显示CDK1、CCNB1、CDC20、CENPE和HIST2H2BE在RMS中的表达与正常对照组相比差异有统计学意义。免疫组化显示CDK1(28/32，87.5%)和CCNB1(26/32，81.25%)在RMS中的表达显著高于正常对照组(1/9，11.1%；0/9，0%)。结论这些HUB基因，尤其是CDK1和CCNB1基因可能是RMS的潜在诊断生物标志物，为RMS的发病机制提供了新的视角。
BackgroundRhabdomyosarcoma (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.MethodsThe 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).ResultsA 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.ConclusionsThese 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.