Background
Laryngeal squamous cell carcinoma (LSCC) is the second most aggressive head and neck squamous cell carcinoma. Although much work has been done to optimize its treatment, patients with LSCC still have poor prognosis. Therefore, figuring out differentially expressed genes (DEGs) contained in the progression of LSCC and employing them as potential therapeutic targets or biomarkers for LSCC is extremely meaningful.
Methods
Overlapping DEGs were screened from two standalone Gene Expression Omnibus datasets, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed. By applying STRING and Cytoscape, a protein–protein network was built, and module analysis was carried out. The hub genes were selected by maximal clique centrality with the CytoHubba plugin of Cytoscape. UALCAN and GEPIA data were examined to validate the gene expression findings. Moreover, the connection of the hub genes with LSCC patient overall survival was studied employing The Cancer Genome Atlas. Then, western blot, qRT-PCR, CCK-8, wound healing and transwell assays were bring to use for further verify the key genes.
Results
A total of 235 DEGs were recorded, including 83 upregulated and 152 downregulated genes. A total of nine hub genes that displayed a high degree of connectivity were selected. UALCAN and GEPIA databases verified that these genes were highly expressed in LSCC tissues. High expression of the SPP1, SERPINE1 and Matrix metalloproteinases 1 (MMP1) genes was connected to worse prognosis in patients with LSCC, according to the GEPIA online tool. Western blot and qRT-PCR testify SPP1, SERPINE1 and MMP1 were upregulated in LSCC cells. Inhibition of SPP1, SERPINE1 and MMP1 suppressed cell proliferation, invasion and migration.
Conclusion
The work here identified effective and reliable diagnostic and prognostic molecular biomarkers by unified bioinformatics analysis and experimental verification, indicating novel and necessary therapeutic targets for LSCC.
The main obstacle to the treatment of nasopharyngeal carcinoma (NPC) is metastasis. Long non‐coding RNAs (lncRNAs) and microRNAs (miRNAs) are highly involved in the progression of NPC. In this study, we aimed to explore the regulatory role of lncRNA P73 antisense RNA 1 T (TP73‐AS1) and miR‐495 in migration and invasion of NPC cells. The expression levels of TP73‐AS1, miR‐495, and junctional adhesion molecule A (JAM‐A) in NPC tissue samples and cell lines were examined by quantitative real‐time PCR (qRT‐PCR) and/or Western blot. NPC cells were transfected with vectors overexpressing TP73‐AS1, short hairpin RNA (shRNA) against TP73‐AS1, shRNA against JAM‐A, miR‐495 mimics, miR‐495 inhibitor, and their corresponding negative controls as designated. The MTT assay, cell migration assay, and transwell assay were performed to detect cell viability, migration, and invasion, respectively. Dual‐luciferase reporter assay was performed to confirm the binding of TP73‐AS1 and miR‐495, and miR‐495 and JAM‐A. TP73‐AS1 and JAM‐A were significantly upregulated while miR‐495 was markedly downregulated in NPC tissues and cell lines compared to normal controls. The overexpression of TP73‐AS1 promoted migration and invasion of NPC cell line CNE‐2. TP73‐AS1 targeted miR‐495 and negatively regulated its expression. TP73‐AS1 upregulated the expression of JAM‐A through miR‐495. TP73‐AS1 mediated migration and invasion of CNE‐2 cells via upregulating JAM‐A. LncRNA TP73‐AS1, miR‐495, and JAM‐A are involved in migration and invasion of NPC cells. The TP73‐AS1/miR‐495/JAM‐A axis may serve as a therapeutic target for the treatment of NPC.
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