EVs Characteristics and Differential miRNA Autophagy Pathway Target Genes in Different Breast Cancer Cell Lines
Objective To study the EVs characteristics and the number and function of miRNA in different breast canc-er cell lines.Methods Normal breast epithelial cells HMEC and breast cancer cell lines MDA-MB-231 and MCF-7 were cul-tured using serum-free medium.The size and content distribution of extracellular vesicles were analyzed by nanoparticle particle size analysis,and the morphology of extracellular vesicles was observed by projection electron microscopy.Extracellular vesicle membrane protein marker CD63 was detected by western bolt.Differential miRNA in extracellular vesicles was analyzed by tran-scriptome sequencing,and differential miRNA target genes were analyzed by GO and KEGG.Results Multi-modal particle size analysis showed that the particle size distribution and content of extracellular vesicles of HMEC were different from MDA-MB-231 and MCF-7.Extracellular vesicles with typical characteristics and the expression of CD63 were detected in all three cell lines.There were 613,333 and 698 known mirnas in extracellular vesicles of HMEC,MCF-7 and MDA-MB-231,respectively.Compared with HMEC,108 and 86 mirnas were significantly up-regulated and down-regulated in extracellular vesicles of MCF-7,and 135 and 246 mirnas were significantly up-regulated and down-regulated in extracellular vesicles of MDA-MB-231,respectively.GO and KEGG enrichment analysis showed that the target genes of different mirnas were involved in multiple biological functions,especial-ly autophagy.Conclusion The diameter,quantity distribution and miRNA content of extracellular vesicles of different breast cancer cell lines are different from those of normal breast cancer epithelial cells,and there are also differences among different breast cancer cell lines.The different miRNA in extracellular vesicles of breast cancer cell lines may have different effects on the autophagy level of the vesicles themselves or their target cells,and the mechanism needs to be further studied.
Breast cancerExtracellular vesiclesNanoparticle size analysisMircoRNAAutophagy
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