LINC02154 Promotes Proliferation,Invasion And Metastasis Of Head And Neck Squamous Cell Carcinoma

ObjectiveHead and neck squamous cell carcinoma(HNSCC)accounts for more than 90%of head and neck carcinoma,with more than 600,000 new cases each year,posing a great threat to human health.At present,the main clinical treatment for patients with HNSCC is a combination of surgery,radiotherapy,and chemotherapy.Although diagnosis and treatment are constantly updated and improved,the five-year survival rate of patients with HNSCC has not been significantly improved.One of the important reasons is the early metastasis of tumors.Studies have shown that more than 60%of patients have metastasis at the time of treatment.Once cervical lymph node metastasis occurs,the five-year survival rate is less than 50%,while the five-year survival rate for distant metastasis is only about 20%.Therefore,it is possible to improve the prognosis of patients with HNSCC by finding specific and sensitive biomarkers for predicting metastasis of HNSCC and taking effective prevention and control measures.LncRNAs is a type of non-coding RNA that is more than 200 nt in length and does not encode protein.At first,it was regarded as the "noise" of genome transcription and did not have biological functions.With the development of genomics,more and more evidences show that LncRNAs can regulate gene expression at the genetic,transcription and post-transcriptional levels,and participate in multiple biological processes such as tumor proliferation,apoptosis,invasion and metastasis.Since LncRNAs can be easily detected in tissues and serum,it is considered to be an emerging marker for tumor diagnosis and prognosis,and may even be a biological target for the new therapeutic methods.In this study,we aim to find LncRNA which is closely related to HNSCC,and conduct a preliminary study on its role in the proliferation,apoptosis,invasion and metastasis of HNSCC in vivo/vitro to explore whether it could be a potential biomarker for HNSCC.Methods1.Download the.clinical data and gene expression data of 111 patients with laryngeal squamous cell carcinoma and 12 patients with normal laryngeal tissue through the Cancer Genome Atlas database(TCGA,https://cancergenome.nih.gov/),edgeR software analyzes differentially expressed genes,LncRNA closely related to the prognosis of laryngeal cancer was obtained through single-gene screening and multifactor cox regression analysis,and a prognostic model of laryngeal cancer based on LncRNA was constructed.2.Quantitative real-time PCR are used to detect the expression level of target LncRNA in head and neck squamous cell carcinoma and normal tissue samples and head and neck squamous cell carcinoma cells.3.Fluorescence in situ hybridization(FISH)detects the subcellular localization of the target LncRNA in head and neck squamous cell carcinoma cells.4.CCK-8,clone assay and cell cycle analysis are done in head and neck squamous cell carcinoma cells after overexpression or knockdown of LINC02154,to detect the growth and survival ability in vitro.5.Flow cytometry are done in head and neck squamous cell carcinoma cells after overexpression or knockdown of LINC02154,to detect the anti-apoptotic ability.6.The Transwell assay are done in head and neck squamous cell carcinoma cells after overexpression or knockdown of LINC02154,to detect the migration and invasion ability.7.qRT-PCR and immunofluorescence are done in head and neck squamous cell carcinoma cells after overexpression or knockdown of LINC02154,to detect the RNA and protein levels of epithelial-mesenchymal transition(EMT)markers.8.The subcutaneous xenograft tumor model of head and neck squamous cell carcinoma in nude mice was established to observe the effect of LINC02154 overexpression or silence on tumor.Results1.LINC02154 was selected as the target gene according to the standard of significant differential expression with the same trend in the database of head and neck squamous cell carcinoma tissue samples.2.The expression of LINC02154 in head and neck squamous cell carcinoma tissues is higher than adjacent tissues,which is positively correlated with clinical stage.There are different expression levels in head and neck squamous cell carcinoma cells,and the expression level of FaDu is higher than AMC-HN-8.4.LINC02154 can improve the growth activity and survival of head and neck squamous cell carcinoma cells,but has no significant effect on cell cycle and antiapoptosis ability.5.LINC02154 can promote the migration and invasion of head and neck squamous carcinoma cells in the Transwell assay.6.qRT-PCR and immunofluorescence found that after overexpression of LINC02154,the expression of epithelial markers E-cadherin decreased,and the expression of mesenchymal markers N-cadherin and Vimentin increased;after silence the expression of LINC02154,the results were opposite to the above trend.7.LINC02154 can promote tumor growth in BALB/C nude mouse.ConclusionsIn this study,we established a 9-LncRNA-based laryngeal squamous carcinoma prognosis model,in which LINC02154 can regulate the growth,migration,invasion,and epithelial-mesenchymal transition of head and neck squamous carcinoma cells,and is expected to become a biomarker for HNSCC diagnosis and prediction of early metastasis.