The Expression And Preliminary Function Study Of LncRNA MANCR In Oral Squamous Cell Carcinoma

Objective Oral squamous cell carcinoma(OSCC)is a kind of head and neck malignant tumor with high recurrence rate and lymph node metastasis rate that seriously threatens human health.Long non-coding RNA(lncRNA)MANCR plays an important regulatory role in the progression of various cancers,but its function in OSCC was largely unclear.The purpose of this study is to explore the expression level of MANCR in OSCC and its influence on OSCC biological behavior,as well as the potential clinical significance and possible function of MANCR,so as to lay a theoretical foundation for lncRNA clinical application translation and targeted therapy of OSCC.Methods In this study,the expression of MANCR in OSCC was determined by the results of previous gene chip,combined with cancer genome map(TCGA)and gene expression database(GEO),and its expression level in OSCC tissues and cells was verified by real-time quantitative PCR(RT-qPCR).Then,the relationship between MANCR and clinicopathological parameters in OSCC tissues was analyzed,and the correlation between MANCR and prognosis of OSCC patients was analyzed by Kaplan-Meier survival curve.Furthermore,the expression level of MANCR was silenced in SCC9 and Cal27 by cell transfection,and the effects of MANCR on proliferation,invasion,migration and apoptosis of SCC9 and Cal27 were investigated by CCK8 experiment,Transwell and flow cytometry,respectively.Finally,the potential role of MANCR in OSCC was preliminarily explored through single gene difference analysis,Gene Oncology(GO)and enrichment analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG).Results1.Biological information analysis showed that MANCR is upregulated in OSCC,and the high expression of MANCR is significantly correlated to the low disease-specific survival(DSS)and overall survival(OS).Meanwhile,upregulation of MANCR is closely related to the low OS of patients with lymph node metastasis,G3/G4 histological grade,T4 stage and clinical stage Ⅳ.Further RT-qPCR results showed that MANCR was highly expressed in OSCC tissues(P < 0.05),and the high expression level of MANCR was closely related to cervical lymph node metastasis and T3/T4 stage of OSCC patients(P< 0.05).2.The results of RT-qPCR showed that MANCR was upregulated in SCC9,Cal27 and HN30 cells compared with normal oral keratinocytes(Hok)(P<0.05).CCK8 assay exhibited the proliferation of SCC9 and Cal27 cells were significantly inhibited after silencing MANCR expression(P < 0.05).Transwell results showed that the invasion and migration of SCC9 and Cal27 cells were markedly reduced after silencing MANCR expression(P< 0.05).The results of flow cytometry revealed that the apoptosis levels of both SCC9 and Cal27 cells were increased after silencing MANCR(P <0.05).3.Single gene difference analysis showed that 214 differentially expressed genes(DEGs)were closely related to MANCR expression(P < 0.05),including 19 high-expression genes and 205 low-expression genes(P <0.01).Furthermore,GO analysis of these DEGs revealed that the high expression of MANCR was significantly related to axonogenesis,neuronal cell body and receptor ligand activity.The results of KEGG analysis exhibited that high expression of MANCR is closely related to neuroactive ligand-receptor interaction,Ras signaling pathway,cell adhesion molecules,the high expression of extracellular matrix and ECM-receptor interaction.Conclusion1.MANCR is highly expressed in OSCC,suggesting that MANCR can function as an oncogene,and the high expression of MANCR is significantly related to the low survival rate of OSCC patients,T3/T4 clinical stage and lymph node metastasis,which indicating that MANCR may probably act as a potential biological marker for evaluating the prognosis of OSCC.2.The downregulation of MANCR can significantly inhibit the proliferation,invasion and migration of OSCC cells,while promote the level of apoptosis,suggesting that MANCR may play a vital role in the initiation and development of OSCC.3.MANCR may affect the initiation and development of OSCC by influencing tumor microenvironment and participating in the process of peripheral nerve regulation.