Construction And Analysis Of Hsa-miR-145-5p-Centered Competing Endogenous RNA Network In Laryngeal Squamous Cell Carcinoma

Objective:The Laryngeal Squamous Cell Carcinoma,referred to as LSCC,ranks the second place in head and neck malignancies,and is highly prevalent in northern provinces of China,with a slight increase in recent years.LSCC has malignant biological behavior that is easy to invade and metastasize,especially cervical lymph node metastasis,and the prognosis of patients is poor.To gradually analyze the gene network regulation of malignant proliferation,invasion and metastasis of laryngeal squamous cell carcinoma is of great significance for us to further understand the mechanism of the occurrence and development of laryngeal squamous cell carcinoma from the perspective of transcriptomics.Previous studies(including ours)have shown that hsa-miR-145-5p plays a role as a tumor suppressor gene in a variety of tumors,including LSCC,and our previous studies have also shown that hsa-miR-145-5p is dysregulated in laryngeal squamous cell carcinoma and is correlated with prognosis.However,the regulation of hsa-miR-145-5p centered gene network is still unclear,which is of great significance for further study of its dysregulated expression and downstream effective targets.This study aims to provide a new idea for further exploring the molecular mechanism of miR-145-5p in inhibiting the malignant progression of laryngeal squamous cell carcinoma from the perspective of the regulatory network of competitive endogenousRNA(ceRNA).Methods:The laryngeal squamous cell carcinoma cells were divided into two groups for culture,and the miR-145-5p overexpression mimics and the control group(NC)mimics were transfected,respectively.RNA was extracted 48 h after transfection(3 samples were repeated in each group),lncRNA microarray(including lncRNA,circRNA and mRNA probes)and miRNA microarray were made,and the original data were obtained by scanning the arrays with Agilent Scanner.The original data were obtained by scanning microarray,and then differentially expressed mRNA,lncRNA,circRNA and miRNA were screened based on the p value<0.05 and fold change >2.The target genes of differentially expressed miRNAs were predicted and functional analysis was performed using the database.The GO biological process enrichment analysis and KEGG pathway enrichment analysis of differentially expressed mRNAs were performed using biological information annotation database DAVID.A regulation network of Competing EndogenousRNA(ceRNAs)centered on miR-145-5p was constructed by bioinformatics technology.Results:According to the microarray results,26 differentially expressed microRNAs(DEmiRNA),248 differentially expressed messengerRNAs(DEmRNA),1118 differentially expressed long non-codingRNAs(DElncRNAs)and 382 differentially expressed circRNAs(DEcircRNAs)were screened out from the laryngeal squamous cell carcinoma cells overexpressed with miR-145-5p.Target genes of differentially expressed miRNAs were predicted,and a total of 675 target genes were obtained.GO function enrichment analysis showed that the differentially expressed genes were mainly related to cell adhesion,and pathway analysis showed that the differentially expressed genes were mainly enriched in MAPK and FOXO signaling pathways.The up-regulated mRNA was involved in the ABC transporter pathway and the down-regulated mRNA was involved in the olfactory transduction pathway.In addition,149 ceRNA pairs were estimated to be associated with apoptosis,Wnt pathway,and metabolic pathway.The q PCR results confirmed that miR-145-5p affected the expressions of lncRNA,miRNA,mRNA and circRNA in LSCC cells.Conclusion:In laryngeal squamous cell carcinoma,hsa-miR-145-5p regulates and forms a complex gene network through ceRNA,and may play an important role in tumor suppression through its mediated important functional molecular pathways.For example,Wnt2B-miR-145-5p-nonhs AT127539.2,CASP10-miR-145-5p-nonhs AT127539.2,CASP10-miR-145-5p-circ000351 and TPO-miR-145-5p-circ 0003519 may play an important biological function in inhibiting laryngeal squamous cell carcinoma,and this provides a new idea for further research on the tumor suppressive function of hsa-miR-145-5p.