| Objective: Try to analysis the miRNA expression profile and screen for specific molecular markers for ovarian immature teratoma in children.Methods: A total of 20 children’s ovarian tumor tissue samples were collected,of which 18 were formalin-fixed paraffin-embedded tumour tissue,including 2 cases of ovarian cyst,3 cases of mature teratoma,6cases of immature teratoma,3 cases of yolk sac tumor,3 cases of juvenile granulosa cell tumor,and 1 case of dysgerminoma;2 were fresh frozen tissue samples,including one mature teratoma and one yolk sac tumor.Differentially expressed miRNAs in samples were screened by Agilent gene chip,which were divided into three groups for statistical analysis.The most important one is the comparation between immature teratomas and the other ovarian tumors(ovarian cyst,mature teratoma,yolk sac tumor,juvenile granulosa cell tumor,dysgerminoma).Differential miRNA between ovarian benign and malignant tumors,between different pathological grade immature teratomas were also analyzed.Then target gene prediction,GO(Gene Ontology)analysis,KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis,miRNA-gene-pathway analysis were performed in miRWalk,miRDB,and DAVID databases to obtains its biological information.Find the miRNAs related to the pathogenesis,diagnosis,treatment,and prognosis of immature teratoma in children.Results:1.Gene microarray screening for differential miRNA: 44 differential miRNA between the benign and malignant ovarian tumor groups were screened.13 different miRNA are specific for immature teratomas(compared with the other 5 types)and 18 miRNA differences among different pathological grades of immature teratoma.2.Bioinformatics analysis:1)The important difference miRNAs between benign and malignant ovarian tumors are miR-106b-5p,miR-20a-5p and miR-20b-5p.The target genes of different miRNAs are mainly involved in transcription and transcriptional regulation(protein binding,RNA polymerase Ⅱ core promoter proximal region sequence-specific DNA binding,transcription factor activity,protein phosphatase regulator activity).Pathway in cancer are the key signal pathway,axon guidance and Fox O signaling pathway are also involved.2)The target gene function of specific miRNA of ovarian immature teratoma is mainly enriched in transcription and transcriptional regulation,including transcription factor activity,RNA polymerase Ⅱ transcription factor activity,sequence-specific DNA binding and protein binding.FOXO1 and AKT3 are involved in the regulation of long-life regulatory pathway,glutamatergic synapse and Circadian entrainment.Vital miRNAs include miR-129-5p,miR-124-3p,miR-7-5p and miR-4698.3)The differently expressed miRNAs in different pathological grades(grade Ⅰ vs grade Ⅱ)of ovarian immature teratomas are related to the development of nervous system.Mi R-4673,miR-378 h and miR-6857-5p are specifically high expressed in grade Ⅰ.Related target genes include MAPK gene family,VEGFA and HGF,which regulate the MAPK signaling pathway,c AMP signaling pathway and neurotrophic signaling pathway.Conclusion:1.The miRNA expression profile of immature teratoma in children for the first time through a microarray analysis of formalin-fixed paraffin-embedded samples and 13 specific miRNAs were obtained.Important miRNAs were found including miR-129-5p,miR-124-3p,miR-7-5p,miR-4698,may be biomarkers for distinguishing ovarian immature teratoma from other ovarian tumors,and laid the foundation for subsequent experiments in blood and fresh tissue samples;2.We explored the function and signaling pathways of relevant target genes of specific miRNA in ovarian immature teratoma in children by bioinformatics analysis,and established the mirna-gene-pathway network,screening out vital target genes and miRNAs;3.Differentially expressed miRNAs were also discovered in different pathology grades of ovarian immature teratomas in children,and these miRNAs are related to the development and differentiation of nervous system,which may be a more objective panel for grade diagnosis in pathology. |
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