Multidrug Resistance-related CeRNAs In Epithelial Ovarian Cancer

Chapter Ⅰ Bioinformatics analysis of drug-resistant ce RNAs in epithelial ovarian cancerObjective: Multidrug resistance in ovarian cancer is a major obstacle to its treatment.In recent years,Lnc RNA has become a hot topic in cancer research.However,the Lnc RNA ce RNA(Lnc RNA-mi RNA-m RNA)has less regulatory mechanisms and has many drugs in ovarian cancer.The study of drug-resistant ce RNAs has not been reported in other research groups.The purpose of this study was to dig out long-chain non-coding competitive endogenous RNA(ce RNA)related to resistance to epithelial ovarian cancer through bioinformatics methods.RNA.METHODS: 25 Lnc RNAs related to epithelial ovarian cancer were retrieved from the PUBMED/MEDLINE database.DIANA-Lnc Base v2,mi Rcode,mi RTar Base,COREMINE,DAVID,mi RSponge,and other related databases were used to predict the possible ce RNA relationship between lnc RNA-mi RNA-m RNA..In the resistant and sensitive tissues of ovarian cancer,Lnc RNA that can be used as ce RNA was verified by QRT-PCR.Finally,Graphpad Prism5 and SPSS19.0 software were used for statistical analysis.Results: Twenty-nine Lnc RNAs located in the cytoplasm wereselected from 25 long-chain noncoding RNAs involved in the development of ovarian cancer.After further bioinformatics analysis and text mining,two Lnc RNAs,GAS5 and HOTAIR,were predicted to serve as competitive endogenous RNAs.(ce RNA),GAS5 and mi RNA-139-5P,mi RNA-24-3P and m RNA(BRCA1,TP53,EGFR,BCL2,FOS,HRAS)compose the ce RNA regulatory network,HOTAIR and mi RNA-30a-5P and m RNA(PIK3R2,TP53(VIM)constitutes a ce RNA regulatory network.Among them,EGFR,BCL2,FOS,and HRAS participate in MAPK signaling pathways,adhesions,pathways in cancer,and other up-regulation pathways,while BRCA1,TP53,PIK3R2,and VIM participate in P53 signaling pathways,and these 9 m RNAs interact with GAS5 and HOTAIR through COREMINE analysis.There is a strong correlation.QRT-PCR results showed that compared with the sensitive group,GAS5 was2.13±1.55-fold up-regulated in the drug-resistant group and 3.39±1.89-fold higher in the HOTAIR group,with a statistically significant difference(P<0.05).The area under the ROC curve(AUC)of GAS5 expression was 0.678,and the area under the ROC curve(AUC)of HOTAIR expression was 0.863.Using Logistic Binary Regression Model to analyze GAS5 and HOTAIR,the area under the ROC curve was 0.871,and the difference was significant(P = 0.000).The COX proportional hazards regression model showed that the relative expression of HOTAIR in tissues was an independent risk factor for PFS and OS in epithelial ovarian cancer,and the relative expression of GAS5 in tissues was an independent factor affecting OS.Conclusion: GAS5 and HOTAIR may be used as ce RNAs to regulate target genes.They have a certain diagnostic efficacy for multidrug resistance of ovarian cancer;HOTAIR has better diagnostic performance than GAS5.Compared with single-indicator diagnosis,combined diagnosis improves the diagnostic efficiency and may provide new ideas for theearly diagnosis of ovarian cancer.Follow-up studies should be conducted for further experiments and clinical double verification.Chapter Ⅱ Prediction and Validation of ce RNA Function in Multidrug Resistance of Epithelial Ovarian CancerObjective: Epithelial ovarian cancer is one of the most common tumors in the female reproductive system.It seriously harms human health and its resistance has always been the focus and difficulty of many scholars.For a long time in the past,m RNA,mi RNA and other studies in epithelial ovarian cancer have been countless,but have not yet significantly reduced their mortality.In recent years,Lnc RNA and ce RNA regulatory mechanisms have attracted more and more attention from researchers.From the analysis of Chapters 1 and 2,we found that only a very few of these Lnc RNA biological functions are involved in epithelial ovarian cancer.verification.At the same time,we analyzed the efficacy of GAS5 and HOTAIR as ce RNAs in the diagnosis of drug-resistant epithelial ovarian cancer through bioinformatics methods.Therefore,we integrated the results of the previous study on m RNA,mi RNA,and Lnc RNA microarrays to obtain epitheliality.The ce RNA network associated with drug resistance in ovarian cancer is expected to provide a long-chain non-coding RNA that can be used as a competitive endogenous RNA(ce RNA)associated with resistance to epithelial ovarian cancer,and to establish a diagnostic model.Combined with the statistical results in Chapter 2,a diagnostic model with high diagnostic efficiency was obtained.Methods: Integrated pre-chip technology was used to screen out the differentially expressed mi RNAs(42 up-regulated,20down-regulated),m RNA(678 up-regulated,308 down-regulated),Lnc RNA(759)in patients with resistant ovarian cancer and sensitive patients.Up-regulation,345 down-regulations;construction of ce RNA network,screening of ce RNA networks associated with mi R-429,mi R-382-5p,and mi R-200b-3p,prediction of possible regulatory mechanisms in ce RNA networks through KEGG pathway enrichment and GO annotation.Using the Lnc RNA in the ce RNA network differential table and the Lnc RNA up-regulated and down-regulated Lnc RNA for QRT-PCR validation,large-sample QRT-PCR validation was performed to verify Lnc RNA consistent with microarray results.Spearman correlation analysis was performed in combination with clinical pathology data.Single-indicator ROC curve,combined with Logistic binary regression model to build joint predictors,K-M single factor survival analysis,COX multi-factor regression model to derive independent risk factors affecting prognosis,and combined with the statistical analysis results of the second chapter to analyze the best combination Diagnostic model.RESULTS: QRT-PCR technique was used to verify the 5 Lnc RNAs that can be used as ce RN in the microarray results:NONOSAT059750,NONSAT129265,NONSAT128169,ENST00000529924,TCONS-12-00003421 and Lnc RNA-NONSAT048424,TCONS-12-00001237 And up-regulation of Lnc RNA NONHSAT116187 expression of a total of 8Lnc RNA,found that in addition to Lnc RNA NONHSAT116187,the verification results of the remaining 7 Lnc RNA and the chip results in the same trend,confirming that the chip is more reliable.Seven Lnc RNAs with large sample validation and consistent chip expression trend results were analyzed: Comparedto the sensitive group,NONSAT059750,ENST00000529924 were up-regulated by 7.62-fold and 2.63-fold in the drug-resistant group,respectively;NONAST129265,NONSAT128169,TCONS-12-00003421,NONOSAT048424,TCONS-12-00001237 were down-regulated by 0.59-fold,0.81-fold,0.51-fold,0.74-fold,and 0.46-fold in the drug-resistant group,respectively,and the difference was statistically significant(P<0.05).According to the expression levels of Lnc RNAs in the resistant group and the sensitive group of epithelial ovarian cancer,the ROC diagnostic curve was drawn.The area under the ROC curve ANOS of NONSAT059750 was 0.949,and the diagnostic efficiency was the best.Combined with Logistic Binary Regression Model to establish a multi-index joint diagnosis of multidrug resistance in epithelial ovarian cancer and draw different ROC curves of the combined factors,combined with the results of the second chapter,obtained by: NONSAT129265+NONSAT128169+ENST00000529924+TCONS-12-00003421+ The area under the curve of the combined predictors composed of GAS5+HOTAI was 0.955.The COX multivariate risk proportional regression model suggested that the relative expression of NONSAT059750 and the size of residual tumor were independent factors influencing PFS.The relative expression of NONSAT059750 and celiac metastasis were independent factors affecting OS.Conclusion: 1.Based on pre-chip data analysis,mi R-429,mi R-382-5p,and mi R-200b-3p related ce RNA(Lnc RNA-mi RNA-m RNA)regulatory networks were constructed.Through the analysis of related KEGG pathways,the ce RNA regulatory network plays a role in multiple signaling pathways.The Lnc RNAs in the screened ce RNA network were verified by QRT-PCR.The results of the preliminary experiments were consistent with the previous data analysis results.3.The combination of NONSAT129265+NONSAT128169+ENST00000529924+TCONS-12-00003421+GAS5+HOTAIR has the best diagnostic performance;Lnc RNANONSAT059750 may be a biomarker for the diagnosis and prognosis of epithelial ovarian cancer.The ce RNA regulatory network may have guiding significance in improving the prognosis of patients with multidrug resistance in epithelial ovarian cancer.The specific regulatory mechanisms need to undergo further experimental research later.Chapter Ⅲ Progress of long non-coding RNA in epithelial ovarian cancerOvarian cancer is the most deadly tumor of the female reproductive system worldwide,and its development involves a variety of genetic and environmental factors.Various studies have identified suitable biomarkers to facilitate early detection,prognostic evaluation,and evaluation of therapeutic responses.This article outlines the role of long-chain non-coding RNAs(lnc RNAs)in the development of ovarian cancer and its potential use as a prognostic factor for ovarian cancer.We searched the MEDLINE / Pub Med database for literature using keywords(including Ovarian cancer,Lnc RNA or long noncoding RNA).We found 25 differentially expressed lnc RNAs that have been validated in ovarian cancer,and some of them have defined their specific mechanisms in ovarian cancer.In this article,we summarize the research progress of Lnc RNAs in epithelial ovarian cancer and find that some lnc RNAs are related to patientsurvival,metastatic potential and treatment response.We look forward to summarizing the results of previous studies and exploring the possibility that lnc RNAs can be used as predictors of drug-resistant predictors of epithelial ovarian cancer through certain mechanisms.