Breast cancer derived exosomes promoted angiogenesis of endothelial cells in microenvironment via circHIPK3/miR-124-3p/MTDH axis

Abstract

Circular RNAs (circRNAs) are important contents in exosomes, which can regulate peripheral cell functions, thus influencing the tumor microenvironment. This work investigated the mechanisms underlying the angiogenesis in peripheral human endothelial cells (ECs) mediated by the breast cancer (BC) cells derived exosomal circRNAs and aimed to explore the biomarkers for the anti-angiogenesis therapy for BC.The BC cell derived exosomes were extracted and the expression level and the circular formation of HIPK3 enclosed was determined. To examine the impact of this exosomal circRNA on ECs, cell viability and tube formation were determined in recipient cells co-cultured with exosomes or transfected with circHIPK3 and the related controls. Target microRNAs (miRNAs) for circHIPK3 and target genes for miRNAs were predicted and confirmed by multiple assays like dual luciferase reporter assay, western blot, and qPCR assays. The existence of the circHIPK3/miR-124-3p/MTDH axis were further confirmed with rescue experiment in mice xenograft model.HIPK3s were mainly in forms of circRNAs and were highly expressed in the BC cell derived exosomes, which could be absorbed by the recipient ECs. The cell viability and angiogenesis in ECs were enhanced when treated with circHIPK3s and decreased when treated with circHIPK3-si. Furthermore, MTDH was proved to be the responsible gene in this process which was regulated by miR-124-3p, the local miRNA sponged by the exosomal circHIPK3.circHIPK3 enclosed in the BC cell-derived exosomes enhanced MTDH expression in the endothelial cell by sponging miR-124-3p, favoring the tube formation in ECs, which might serve as a therapeutic target for anti-angiogenesis therapy for breast cancer.