Effect of human chorionic trophoblast cells-derived miR-155-containing exosomes on the proliferation and tube-forming capacity of human umbilical vein endothelial cells
Objective To construct miR-155-containing exosomes derived from human chorionic trophoblast cells,and to investigate the effects of miR-155 carried by exosomes on the proliferation and tube-forming ability of human umbilical vein endothelial cells(HUVECs).Methods Fresh aborted tissue with normal development in early pregnancy was collected for primary culture of human chorionic trophoblast cells in Xi'an People's Hospital(Xi'an Fourth Hospital)from January 2023 to April 2023,and exosomes were extracted and collected in the cell supernatant;transmission electron microscopy,nano-particle tracking analysis(NT A),and Western-blot experiments were performed to identify the exosomes.Exosomes containing Exo-miR-155 mimics,Exo-miR-155 inhibitor,and Exo-NC were constructed by drug-loading technology;q-PCR was used to detect the expression of miR-155 in exosomes.The constructed exosomes were then co-incubated with HUVECs respectively,and fluorescence microscopy was used to detect exosome uptake;CCK-8,tube-forming assay were used to detect the proliferation and tube-forming ability of HUVECs cells after exosome uptake.CYR61 and VEGF expression in HUVECs cells were detected by q-PCR and Western-blot.Results Isolated cultured human placental chorionic trophoblast cells were able to significantly express CK-7;exosomes extracted from human placental chorionic trophoblast cell cultures had a typical morphology,particle size and expressed the exosomel characteristic proteins CD9 and CD63;the expression and inhibition effect of miR-155 in the exosomes were significant;compared with HUVECs after uptake of Exo-NC,HUVECs proliferation and tube-forming ability were decreased after uptake of Exo-miR-155 mimics;and the expression of CYR61 mRNA,VEGF mRNA and protein expression were reduced in cells.Conclusion The carriage of miR-155 by trophoblast-derived exosomes can affect the ability of HUVECs to proliferate and form tubes,as well as the expression of angiogenesis-associated proteins CYR61 and VEGF in the cells,which correlates with the abnormal establishment of placental function and maternal systemic vascular endothelial dysfunction in PE.
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