CDK1 and CCNB1 as potential diagnostic markers of rhabdomyosarcoma: validation following bioinformatics analysis

Abstract

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.

Pax3 inhibits Neuro‐2a cells proliferation and neurite outgrowth

“…48 However, bioinformatics analysis exhibited that CCNB1 was significant up-regulated in F I G U R E 7 KEGG analysis of Neuro-2a cells infected by Ad-Pax3. KEGG pathways analysis revealed 'Parkinson disease', 'Protein processing in endoplasmic reticulum' and 'cell cycle' were significant KEGG pathways compared with untreated cells Rhabdomyosarcoma patient, 49 which was not consistent with the finding in Neuro-2a cells overexpressing Pax3. It indicated that CCNB1 may not be the direct target of Pax3.…”

miR-378a-5p inhibits the proliferation of colorectal cancer cells by downregulating CDK1

“…Besides, the overexpression of CDK1 promoted CRC cell proliferation, invasion and migration, indicating the promotive role of CDK1 in CRC development. Some researchers have found that CDK1 and cyclin B1 may be potential diagnostic biomarkers for rhabdomyosarcoma and hepatocellular carcinoma [16,17]. As discovered by Yamamura et al, phosphorylated CDK1 (p-CDK1), p-CDK2, cyclinB 1, and cyclinE 1 levels increase within cholangiocarcinoma tissues; in the meantime, P-CDK1 and cyclin B1 nuclear levels were positively correlated with the clinical stage as well as lymph node metastasis, while the expression of p-CDK 1 is related to poor patient survival [18].…”

Bioinformatic Identification and Validation of Key Genes and Biological Pathways Involved in Hepatitis B Virus-related Hepatocellular Carcinoma

“…Reports indicated that knockdown of CCNB1 regulated by microRNA-144 signi cantly inhibited cell proliferation, migration, and invasion in HCC [31]. Besides, CCNB1 is also a promising biomarker for ER+ breast cancer [32] and rhabdomyosarcoma [33].…”