Preliminary Screening And Identification Of Long Non-coding RNA Related To Laryngeal Cancer Based On High Throughput Sequencing Technology

Background:Laryngeal cancer is a kind of common malignant tumor of the head and neck,accounting for 5.7%~7.6% of the total malignant tumors.The incidence and mortality of laryngeal Cancer are 2.3/100,000 and 1.2/100,000 respectively all over the world,which is a serious threat to human health.However,with the acceleration of urbanization,industrialization and population aging,high smoking rate and increasingly serious environmental pollution,the incidence of laryngeal cancer in China has an increasing trend.At present,the diagnosis of laryngeal cancer is still made by the combination of patients’ clinical manifestations,imaging features,histological examination results and other clinical auxiliary examination methods.The limitations of these traditional methods,to some extent,make the early diagnosis and treatment of laryngeal cancer difficult.The pathogenesis of laryngeal cancer is still unknown.Therefore,it has become a research direction with potential clinical value to find effective tumor markers that can be applied in clinical laryngeal cancer screening and early diagnosis by revealing the molecular mechanism of laryngeal cancer.With the rapid development of gene chip and high-throughput sequencing technology in recent years,transcriptome level sequencing has been pushed to a new level.Studies have found that long non-coding RNA(lncRNA),which accounts for more than 95% of the genome,is widely involved in various processes of the development and change of human tumors through a variety of mechanisms,and has potential clinical research value as a molecular marker for the early diagnosis and prognosis of malignant tumors as well as a target for molecular targeted therapy.Currently,studies on lncRNA related to laryngeal cancer are still in its infancy.Theestablishment of lncRNA expression profile and the exploration of specific lncRNA in the pathogenesis of laryngeal cancer have become a new focus in the field of cell and molecular biology.Objective:Based on the high-throughput sequencing technology,lncRNAs that may be related to the occurrence and development of laryngeal squamous cell carcinoma were screened by comparing the differences of long noncoding RNA between laryngeal squamous cell carcinoma and paracancer tissue,in order to find potential molecular markers related to laryngeal cancer.Research methods:1.LncRNA expression profile related to laryngeal cancer was obtained based on high-throughput sequencing,and the differentially expressed lncRNA was analyzed by bioinformatics method to screen out the differentially expressed lncRNA in laryngeal cancer and adjacent tissues.2.Enrichment of differentially expressed lncRNA and its related genes in the signaling pathway.6 lncRNAs with significant differences were screened according to the differential expression multiples of lncRNAs.3.Select in November 2017-September 2018 in the second hospital of Jilin university diagnosis of laryngeal cancer and in otolaryngology head and neck surgery and the pathology,the first hospital diagnosed as laryngeal cancer resection patients of laryngeal squamous cell carcinoma of the larynx,beside the corresponding carcinoma specimens,using Reverse transcription-polymerase Chain Reaction(RT-PCR)detection which 8 cases of laryngeal carcinoma tissues,tissue adjacent to carcinoma lncRNA expression in 6 purpose,According to RT-PCR results,two lncRNAs with the most obvious trend of differential expression were selected for quantitative fluorescence qPCR verification in 15 pairs of laryngeal cancer and adjacent tissues.Results:593 lncRNAs were differentially expressed in laryngeal cancer and the corresponding adjacent tissues,among which 312 lncRNAs and 228 lncRNAs weredifferentially expressed at 4 times or more and 8 times or more respectively.6lncRNAs with significant expression differences were selected,namely NONHSAT244419.1,NONHSAT185102.1,NONHSAT176191.1,NONHSAT176190.1,NONHSAT176189.1,NONHSAT149828.1.LncRNA NONHSAT244419.1,NONHSAT176190.1,NONHSAT176189.1,NONHSAT149828.1 were differentially expressed in 15 pairs of laryngeal cancer and adjacent tissues and normal tissues,and the results were homogeneous with high-throughput sequencing results.Conclusion:1.LncRNA expression profile of laryngeal cancer was preliminarily established based on high-throughput sequencing technology,which showed a large number of differences compared with the paracancer tissues.2.The application of high-throughput sequencing technology to the sequencing of laryngeal cancer and adjacent tissues can provide a large number of tumor related data at the transcriptome level,which is helpful for the discovery of new laryngeal cancer markers and related mechanisms.3.NONHSAT176190.1,NONHSAT176189.1 was differentially expressed in laryngeal cancer and adjacent tissues,and was down-regulated in laryngeal cancer tissues.LncRNA can be used for further verification.