Research On The Effect Of Long Non-coding RNA-CYTOR On The Prognosis Of Patients With Ewing’s Sarcoma

Background and ObjectivesEwing’s Sarcoma(ES)is the second most common primary malignant bone tumor in children and adolescents.It has the characteristics of high incidence,high malignancy,high mortality and disability rate,easy metastasis and easy recurrence.The 5-year survival rate of patients with locally unmetastasis Ewing’s sarcoma can reach 70%after scientific multi-modal combination therapy,but the 5-year survival rate of patients with early metastasis is less than 30%.The poor prognosis of Ewing’s sarcoma has brought a huge physical,psychological and economic burden to the patient,his family and the whole society.The main reason for this dilemma is that the factors and detailed mechanisms affecting the prognosis and pathological progress of Ewing’s sarcoma have not been fully elucidated.Many recent studies have shown that long-chain non-coding RNA(lnc RNA)can play an indispensable role in the occurrence and development of diseases,especially tumors,as oncogenes or tumor suppressor genes,and affect the clinical prognosis of tumor patients.Among them,lnc RNA-CYTOR has been proved to be highly expressed in gallbladder cancer,gastric cancer,lung cancer and other cancers as an oncogene and affect its clinical prognosis.However,there is no research report on the expression level of CYTOR in Ewing’s sarcoma and the influence of CYTOR on the prognosis of patients with Ewing’s sarcoma.Therefore,the purpose of this study is to explore and verify the expression level of CYTOR in Ewing sarcoma,and to explore the impact of CYTOR on the prognosis of Ewing sarcoma patients,and to try to predict the molecular mechanism of CYTOR affecting the prognosis of Ewing sarcoma patients。And try to predict small molecule drugs that have potential therapeutic effects on Ewing’s sarcoma,and ultimately link basic research with clinical prognosis and treatment,and try to provide a new idea for the clinical transformation of basic experiments.Methods:1.Purchase and culture human Ewing sarcoma cell lines(TC-32,RD-ES)and normal control cell lines(human bone marrow mesenchymal stem cells,HBMSC),and use RT-q PCR to detect the expression level of CYTOR in each cell line;2.Obtain the gene expression data and clinical prognosis related information of56 patients with Ewing’s sarcoma from the International Cancer Genome Consortium(ICGC).And carry out batch calibration and standardization,and then divide them into CYTOR low expression group and CYTOR high expression group according to the expression level of CYTOR;3.Perform comprehensive prognostic analysis(Kaplan-Meier survival analysis,univariate analysis,multivariate analysis and ROC curve)on standardized data to explore the impact of CYTOR on the prognosis of patients with Ewing’s sarcoma;4.Perform gene set enrichment analysis(GSEA)on the standardized data to predict the molecular mechanism of CYTOR affecting the prognosis of patients with Ewing’s sarcoma;5.Perform gene co-expression analysis on the standardized data to predict CYTOR co-expressed genes;6.Perform CMap small molecule drug analysis on the standardized data to predict small molecule drugs that have potential therapeutic effects in the clinical treatment of Ewing’s sarcoma and have a certain impact on the prognosis of patients with Ewing’s sarcoma.Results:1.The results of RT-q PCR suggested: The gene expression level of CYTOR in Ewing sarcoma cells(TC-32,RD-ES)was significantly higher than that of CYTOR in the corresponding normal control cells(HBMSC)(P< 0.0001);2.The results of Survival analysis suggested: In the CYTOR high expression group,the overall survival of patients with Ewing’s sarcoma was significantly lower than the overall survival of the CYTOR low expression group(p=0.022).The results of single and multiple factors analysis suggested: The abnormally high expression of CYTOR was an independent risk factor related to the prognosis of patients with Ewing’s sarcoma(HR>1,p<0.05);The results of ROC curve suggeste: The area under the curve(AUC)of 1 year and 3 years are both greater than 0.7,indicating that the abnormally high expression of CYTOR has a certain evaluation value for the prognosis of Ewing’s sarcoma3.The results of GSEA suggested: In the high expression group of CYTOR,CYTOR is significantly enriched in the following three cancer-related cell signaling pathways-“extracellular matrix receptor interaction cell signaling pathway”,“cytokine-cell Factor receptor interaction cell signaling pathway” and “Ig A production related intestinal immune network cell signaling pathway”;4.The results of gene co-expression analysis suggested: CYTOR is positively correlated with the gene expression of MIR4435-2HG,SH3BGRL3,NEXTIN2,NFKBIE and S100A16(correlation coefficient>0.6,p<0.05);CYTOR is negatively correlated with the gene expression of ZBTB26,RNF20,DCAF7,RBBP4 and MRPL50(correlation coefficient Correlation coefficient<-0.6,p<0.05);5.The results of CMap small molecule drug analysis suggested: Melatonin,Propranolol,Nimesulide and Leflunomide may have the potential to treat Ewing’s sarcoma.Conclusion:1.Long non-coding RNA-CYTOR has abnormally high expression in Ewing’s sarcoma cells and is a potential oncogene;2.The abnormally high expression of CYTOR can be used as an independent risk factor to lead to poor prognosis of patients with Ewing sarcoma,and it has certain predictive value for evaluating the prognosis of patients with Ewing’s sarcoma;3.It is predicted that CYTOR may be involved in the regulation of three cancerrelated cell signaling pathways: extracellular matrix receptor interaction,cytokine-cytokine receptor interaction,and Ig A production related intestinal immune network to promote the malignant process of Ewing’s sarcoma and lead to its poor prognosis,but the specific mechanism remains to be further studied.4.Prediction: Melatonin,Propranolol,Nimesulide and Leflunomide may have potential therapeutic effects on Ewing’s sarcoma,or may affect its prognosis,but the specific mechanism remains to be further research.