Study On The Differential Expression Profiling Of IncRNAs In Radiosensitivity Of Nasopharyngeal Carcinoma

Objective: This study used next-generation sequencing(RNA-seq)technology to obtain differential lncRNA expression profiles of nasopharyngeal carcinoma before and after radiotherapy,to study the mechanism of lncRNA in nasopharyngeal carcinoma,and to find blood-related lncRNAs that are sensitive to nasopharyngeal carcinoma radiotherapy.The markers provide experimental basis for the clinical individualized radiotherapy for nasopharyngeal carcinoma.Methods: Patient with nasopharyngeal carcinoma who received radiotherapy alone was selected as the research object.The peripheral venous blood samples before and after radiotherapy were collected.The RNA was extracted and the purity of total RNA was detected.By using RNA-seq to detect changes in the expression of lncRNAs before and after radiotherapy,the differential expression profiles of lncRNAs before and after radiotherapy for nasopharyngeal carcinoma were obtained.Quantitative Real-time PCR was used to verify differential expression profiles.Sequencing data were analyzed using bioinformatics software,and differentially expressed lncRNAs were analyzed by gene ontology(GO)analysis and pathway enrichment analysis methods to obtain signal pathways related to radiosensitivity of nasopharyngeal carcinoma and possible participation.Cell components,molecular functions,and biological processes.Quantitative Real-time PCR was used to detect the relative expression of lnc-CRTC3-3 in 30 patients with nasopharyngeal carcinoma before radiotherapy,and the relationship between the expression of lnc-CRTC3-3 and clinicopathological parameters of nasopharyngeal carcinoma was analyzed.Use Graphpad Prism 5 software for statistical analysis and plotting.Results: 1.RNA-seq lncRNA test results: There were statistically significant differences in the expression of 1097 lncRNAs before and after radiotherapy(P<0.05,and the absolute value of log2(fold change>0),including 418 upregulated lncRNAs And 679 expressed downregulated lncRNAs.2.Gene Ontology(GO)analysis results: The GO enrichment analysis of differentially expressed targeted genes of lncRNAs results in: the biological process with the highest degree of enrichment is a cellular process,and the most enriched cellular component is the intracellular part,and the most enriched molecular function is the protein binding function.3.KEGG enrichment pathway analysis results: We analyzed differentially expressed targeted genes of lncRNAs for KEGG enrichment and obtained 10 related signaling pathways.One of the more critical pathways is the N-glycan biosynthetic pathway.4.Verification of differential expression profiles using real-time PCR.Two changes in the experiment were selected to significantly upregulate lncRNA(lnc-SURF2-1,lnc-KXD1-1)and downregulate lncRNA(lnc-EPS8L2-3,lnc-CRTC3-3)and detect their expression in blood samples before and after radiotherapy in patients with nasopharyngeal carcinoma.The results confirm that the trend of gene up-regulation and down-regulation is consistent with the sequencing results.5.Expression of lnc-CRTC3-3 in nasopharyngeal carcinoma and its correlation with clinicopathological parameters of nasopharyngeal carcinoma: The expression level of lnc-CRTC3-3 was closely related to the clinical stage(P=0.0125),differentiation(P=0.0031)and radiosensitivity(P=0.0139),and the expression level of lnc-CRTC3-3 in nasopharyngeal carcinoma patients.The patient's gender,age,history of smoking,and lymph node metastasis were not significantly correlated.Conclusion: 1.The differential expression profiles of lncRNAs associated with NPC radiosensitivity were obtained.2.lncRNAs affect the sensitivity of NPC cells to radiotherapy through relevant signaling pathways.3.The expression level of lnc-CRTC3-3 is closely related to clinical stage,differentiation and sensitivity to radiotherapy in patients with nasopharyngeal carcinoma.lnc-CRTC3-3 may be a potential marker for predicting radiosensitivity of nasopharyngeal carcinoma.