Effects of exosomes from HCT116 cells overexpressing miR-193b-3p on proliferation,apoptosis,migration and angiogenesis of HUVEC and their targeted relationship with PAK4
Objective To observe the effects of exosomes from human colorectal cancer(CRC)cells(HCT116)with overexpression of microRNA 193b-3p(miR-193b-3p)on the proliferation,apoptosis,migration and angiogenesis of human umbilical vein endothelial cells(HUVECs),and to analyze the targeted relationship between miR-193b-3p and p21-activated kinase 4(PAK4).Methods Serum from CRC patients and exosomes in the supernatant of HCT116 cells were collected.Transmission electron microscopy and nanoparticle tracking analysis were used to observe and identify exo-somes.RT-qPCR was performed to detect exosomal miR-193b-3p and PAK4 mRNA.HUVECs were taken and grouped.HUVECs in the PBS group were added with 100 µL of PBS,HUVECs in the HCT116-Exo group were added with 100 µL of HCT116 cell exosomes,and HUVECs in the HCT116-Exo+GW4869 group were first treated with GW4869(10 µmol/L)for 30 min and then were added with 100 µL of HCT116 cell exosomes.The proliferation,migration and angiogenesis abili-ties of HUVECs in each group were detected by EdU assay,Transwell assay and tube formation assay.HUVECs in the log-arithmic growth phase were taken and divided into the miR-NC group and the miR-193b-3p group.HUVECs in the miR-NC group were co-cultured with HCT116 cell exosomes transfected with miRNA negative control(miR-NC),and the miR-193b-3p group with HCT116 cell exosomes transfected with miR-193b-3p mimics.The activity,cell cycle,apoptosis and angiogenesis ability of HUVECs in each group were detected by CCK-8 assay,flow cytometry and tube formation as-say.The targeted relationship between miR-193b-3p and PAK4 was detected by the dual-luciferase reporter gene sys-tem.Results The expression of miR-193b-3p in serum and exosomes of CRC patients increased,while the expression of PAK4 mRNA decreased(both P<0.05);exosomes could promote the proliferation,migration,and angiogenesis of HU-VECs(all P<0.05).Overexpression of miR-193b-3p increased the cell viability and angiogenesis,accelerated cell cycle progression,and inhibited the apoptosis of HUVECs(all P<0.05);miR-193b-3p inhibited its luciferase activity by direct-ly targeting PAK4(P<0.05).Conclusions The expression of miR-193b-3p in exosomes of CRC serum and cancer cells increases,and the proliferation,migration and angiogenesis of HUVECs with the addition of HCT116 exosomes are en-hanced.HUVECs added with HCT116 exosomes transfected with miR-193b-3p mimics show increased proliferation,S phase proportion,angiogenesis ability,and decreased G0/G1 phase proportion and apoptosis rate;miR-193b-3p has a tar-geted relationship with PAK4.
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