Expression Of Exosomal MiRNA In Serum Of Breast Cancer Patients And Analysis Their Target Genes Function

Objective The aim of the study was to investigate the expression and clinical significance of Exosomal mi RNA in serum of patients with breast cancer and speculate its possible function mechanism by bioinformatics analysis and assess the potential of these Exosomal mi RNAs as novel therapeutic targets for breast cancer.Methods(1)The serum from 72 cases of breast cancer patients is our research object,including 42 had stage I and II breast cancers,30 had stage ?and ?breast cancers;29 had lymph node metastases,and 26 patients of estrogen,progesterone,and Her2-neu receptor status(ER,PR,and Her2)was negative with breast cancer and 38 healthy volunteers as controls.(2)Determined the presence of Exosomes by Western blot and the morphology of the Exosomes observed by transmission electron microscopy.(3)Detect the expression levels of the above Exosomal mi RNA by RT-PCR.The relationships between clinicopathologic features of breast cancer and the expression levels of Exosomal mi RNAs were analyzed.(3)Download the date of the breast cancer mi RNA expression from The Cancer Genome Atlas database and compare it with our study.(4)The software,including Mi Randa ? mi RBase and Target Scan,were used to predict the target genes of the verified Exosomal mi RNAs,GO and KEGG pathway enrichment analyses for the identified target genes were performed using DAVID database.Results(1)Eosomes exhibited the expected sizewith a size ranging from 30 to 100 nm of a characteristic round or cup-shaped morphology in patient and expressed marker proteins CD63.(2)Exosomal mi R-21 ? mi R-105 ? mi R-182 ? mi R-122 ?mi R-1246?mi R-373 and mi R-222 are higher in breast cacer patients than that in healthy subjects(P?0.05),while exosomal mi R-378 e are lower in breast cancer patients(P?0.05).Exosomalmi R-182?mi R-222?mi R-373?mi R-21 are higher in breast cacer patients with stages?and ?(P?0.05),while Exosomal mi R-122?mi R-105?mi R-1246?mi R-378 ere no significant difference in the expression level of breast cancer patients with different stages.The expression level of Exosomal mi R-122?mi R-222?mi R-373?mi R-21 are higher in breast cancer patients with lymph node metastasis(P?0.05).Exosomal mi R-182?mi R-105?mi R-1246?mi R-378e(P? 0.05)was no significant difference in the expression of lymph node metastasis(P>0.05)The expression level of Exosomal mi R-105?mi R-373?mi R21?mi R-222 are higher in breast cancer patients with Triple negative(P ? 0.05).Exosomal mi R-122?mi R-182?mi R-1246?mi R-378 e was no significant difference in the expression of non-Triple negative(P?0.05).The result of our study,compared with the TCGA database,shows that: Compared with the high expression of Exosome in peripheral blood serum,mi R-21?mi R-105?mi R-182?mi R-122 are higher in breast cancer patients than that in healthy subjects(P?0.05).(4)The target genes of mi R-105,mi R-373,mi R21,mi R-122,and mi R-222 were predicted by Mi Randa,mi RBase and Target Scan software and analyzed GO analysis and KEGG pathway analysis using the DAVID database.GO analysis revealed that the target genes related to mi RNAs was mainly focused on cell cycle regulation,cell proliferation,and negative regulation of cell physiological processes.KEGG pathway analysis revealed that breast cancer Exosomal mi RNA-related genes are mainly involved in tumor signaling pathways,MAPKs signal pathways,PI3K-Akt signaling pathways,Fox O signaling pathways,and glucose metabolism pathways etc,are involved in the development of breast cancer.Among them,the most significantly activated one is MAPK signaling pathway.Conclusion(1)In this study,The serum Exosomes are significantly dysregulated in breast cancer.(2)Exosomal mi R-21?mi R-105?mi R-182?mi R-122?mi R-373 and mi R-222 are significantly upregulated in breast cancer and show closely related to linicopathologic features.(3)GO analysis and KEGG pathway analysis revealed that the target genes might involve with known breast cancer-related biological processes and signaling pathways.In conclusion,these Exosomal mi RNA correlated significantly with breast cancer invasion,metastasis and could be a novel therapeutic target of breast cancer.