MicroRNA-mRNA Different Expression With Multi-drugresistence In Epithelial Ovariancancer

Ovarian cancer accounts for third place in the female reproductive tract cancer incidence,but the mortality rate ranks first.Chemotherapy is an important treatment for epithelial ovariancancer(EOC).But multidrug resistance of EOC is a vital link which lead to Chemotherapyfailure. Present study suggests those molecular mechanisms of multidrug resistance of EOCmight involve intracellular chemotherapeutic drug efflux,DNA damage and repair, abnormalMolecular mechanisms of apoptosis and signal transduction pathway disorders. But has yet tofind a safe and effective drug target for reversal multidrug resistance.This study intends todiversify through the use of research tools miRNA microarray technology and gene chiptechnology, and bioinformatics, from clinical studies to basic research and finally returnclinical strategies.Looking for multi-drug resistance molecular mechanism in ovarian cancerfor finding security effective reversal of multidrug resistance. Objective Construction miRNA exprssion profiles in epithelial ovarian cancer multidrugresistance.Materials and Methods Using Agilent miRNA chip,detect the miRNAs expressionbetween the five cases of epithelial ovarian cancer tissues with5cases of drug-resistant tissueand5sensitive one,5cases of benign ovarian tumors and5cases of normal ovarian tissue.Ttest and Fc differences was be used to find greater different than2-fold up-regulation ordown-regulation in miRNAs. We mined the data of microRNA-target genes via multiplesonline software.We analysised the target gene of up-regulation or down-regulation miRNAsby GO enrichment analysis and Pathway bioinformatics analysis to predict potential targetgenes and signaling pathways involved in the biological functio. And then we validate themicroarray results using QRT-PCR analysis and expand the sample size to detect theexpression of purpose miRNA.And analyze the relationships between miRNA expressionlevels and clinicopathological factors.Results A total of62microRNAs expressed differentially related to EOCchemoresistance,there were42miRNAsincreased in ovarian cancer resistance tissue and20ones decreased.And1391target genes were predicted by up-regulation miRNAs,1231targetgenes were predicted by down-regulation miRNAs.GO analysis showed:miRNAs ofup-regulated predicted target genes are involved in regulation of cellular protein metabolicprocess and regulation of transcription from RNA polymerase II promoter.Down-regulateddifferentially predicted target genes are involved in regulation of transcription, proteolysisinvolved in cellular protein catabolic process, cellular protein catabolic process and otherclass of biological process-es.Pathways pathway analysis is enriched to TGF-beta signalingpathway and Adherens junction a signaling pathway for up-regulated predicted target genes.Enrichment for disease analysis showed that part of the target genes associated with epithelialovarian cancer or ovarian cancer.We screened four upregulated genes of miR-382-5p,miR-1,miR-152and miR-299-5p and one down-regulated miR-200a-3p carried by QRT-PCRmicroarray validation. The result matched with the microarray results,expand clinical sampleobtained consistent results,which miR-200a-3p was downregulated0.31-fold in ovariancancer drugresistant tissue via drugsensitive tissue,the difference was statistically significant (P<0.01);miR-382-5p,miR-152and miR-299-5p were raised3.53times,2.69times and2.70times, the differences were statistically significant (P<0.01),miR-1was raised1.14times,butthe difference was not statistically significant (P>0.05).Conclusion We screened out miRNA expression profiling associated with epithelialovarian cancer multidrug-resistance by miRNA microarray.the result of predicting targetgeneprompted that miRNA can regulate multiple target genes, and a target gene may also beregulated by several of miRNA.Visiblely miRNAregulatory network is very complex.Bioinformatics analysis showed differential expression of miRNA target genes predicted tobroad participation in a number of biological processes,we speculated that these abnormalexpression of miRNA expression is likely through regulatedof target gene take part in cellproliferation and apoptosis related factors,and involved in several signal transductionpathways,may play an important role in tumor growth and cell prognosis.There aesneed forfurther indepth study of these differentially expressed miRNA to identify potential Molecularmechanisms underlying drug resistance in ovarian cancer,as well as clinical diagnosis andtreatment of individual markers, even to effectively reverse the resistance of epithelial ovariancancer treatment provide new ideas. Objective Construction mRNA exprssion profiles in epithelial ovarian cancer multidrugresistance.Materials and Methods Using Agilent lncRNA chip,detect themRNAs expressionbetween the five cases of epithelial ovarian cancer group with5cases of drug-resistant tissueand5sensitive one,5cases of benign ovarian tumors and5cases of normal ovarian tissue.Ttest and Fc differences was be used to find greaterdifferent than2-fold up-regulation ordown-regulationin mRNAs.We analysised the target gene of up-regulation or down-regulationmiRNAs by GO enrichment analysis and Pathway bioinformatics analysis to predict thebiological functions and involved inwhich signaling pathways of the differential expression genes. And then we validatethe microarray results using QRT-PCR analysis and expand thesample size to detect the expression of purpose mRNA.And analyze the relationships betweenmiRNA expression levels and clinicopathological factors.Results A total of855microRNAsexpressed differentially related to EOCchemoresistance,there were598mRNA increased in ovarian cancer resistance tissue and257ones were decreased.Bioinformatics analysis showed that: up-regulation mRNA involved incell adhesion,biological adhesion,regulation of cell prolifera-tion,immune response,bloodvessel development, and other class of biological processes.Down-regulation mRNA is notinvolved in significantly enrichment related biological processes.KEGG Pathway analysisshowed upregulated mRNA related pathways involved in Focal adhesion and ECM-receptorinteraction.We screened four upregulated genes of KIT,HGF,CSF1R,PDGFRB and onedown-regulated ERBB4gene carried by QRT-PCR microarray validation,the results matchedwith the microar-ray results,expand clinical sample obtained consistent results,whichKIT,HGF,CSF1R, PDGFRB genes in EOC drugresistance group were raised3.81times and2.92times2.74,2.03times via drugsensitive group,the difference was statistically significant(P<0.05), while the KIAA1804gene is down-regulated0.42-fold, difference was statisticallysignificance (P<0.05).Conclusions Microarray-based screening out mRNA expression profiles associated withepithelial ovarian cancer multidrug resistance,which enriching the current data known targetgenes associated with drug resistance.Bioinformatics analysis of the differentially expressedmRNA principalinvolved in cell adhesion,biological adhesion,regulation of cell proliferation,immune response, as well as part of the mRNA is enriched in Focal adhesion and ECM-receptor interaction, suggesting that these differentially expressed genes may be involved inepithelial ovarian cancer multi-drug resistant through a variety of ways. ObjectiveConstruction miRNA-mRNA regulatory networks associated with epithelialovarian cancer multidrug resistance.Materials and Methods To screen out the miRNA and mRNA expression profiling ofepithelial ovarian cancer multidrug resistance,we analysised the differences expressionmiRNA and mRNA in expression profiling information using MAGIA meta-analysis software,and select miRNA negatively correlated with mRNA of miRNA/mRNA,build miRNA-mRNAregulatory network diagram associated with epithelial ovarian cancer drug resistance, andverify that the key expression of miRNA and mRNA using QRT-PCR methods.Results Received a total of53pairs of miRNA-miRNA for negative association,including10miRNA and43mRNA.miR-429and miR-381were enriched by many targetgene, corresponding to the number of target genes was16and8,respectively.The mRNA ofERBB4,PRKCE and PPP1R9A were associated with several miRNA. It was confirmed thatmiRNA regulation a multiple target genes, which gene subject to more than one miRNAregulation. Expanded clinical samples QRT-PCR validation results: compared to ovariancancer sensitive group, miR-429, miR-373-5p in ovarian resistance group were down0.35-fold and0.39-fold,the difference was statistically significant (P<0.01); Thecorresponding target genes KANK2, ZEB2and CTSD were raised2.56times,3.15times and1.89times, the difference was statistically significant (P<0.05); miR-381, miR-495andmiR-410in the drug group were raised2.36times,2.46times and2.42times,the differencewas statistically significant (P<0.01); corresponding target genes ERBB4, TIAL1, FGFRL1and MPPED2were down0.46times,0.38times,0.29times and0.43times, the differencewas statistically significant (P<0.05).Conclusion After QRT-PCR validation and text mining, we believe that to buildmiRNA-mRNA regulatory network diagram with epithelial ovarian cancer multidrugresistance associated with more reliable.It was display that the miRNA and mRNA complexregulatory network relationshipsin ovarian cancer resistance tissues,also from the perspectiveof multiple genes and multi-targets mechanisms of epithelial ovarian cancer multidrugresistance.The key miRNAand its negative regulation of mRNA in which specific ways andmeans to be involved in multidrug resistance mechanisms EOC, needs to make furtherin-depth functional studiesin order to find a really reliable way that can reversal the epithelialovarian cancer MDR,and happened to better guide clinical Chemical therapy and a effectiveprevention avoid multidrug resistance in ovarian cancer patient,expec-ting improve theprognosis and quality of life for EOC patients.