Expression of miR-181a-5p in serum of ACS patients and its regulatory effects on proliferation and migration of human HCASMC cells,and its targeting relationship with ADAMTS1
Objective To investigate the expression of microRNA-181a-5p(miR-181a-5p)in the serum of patients with acute coronary syndrome(ACS)and its regulatory effects on the proliferation and migration of human coronary artery smooth muscle cells(HCASMC),and to explore the targeting relationship between miR-181a-5p and a disintegrin and me-talloproteinasewith thrombospondin motifs 1(ADAMTS1).Methods Peripheral venous blood was collected from 100 patients with ACS[ACS group,including 55 cases of acute myocardial infarction(AMI)and 44 cases of unstable angina pectoris(UAP)]and 40 cases of negative coronary angiography(control group).Real-time quantitative PCR was used to detect the serum miR-181a-5p in the two groups.HCASMC in the logarithmic growth phase were divided into groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,which were transfected with miR-181a-5p mimics,mimics-NC,miR-181a-5p inhibitor,and inhibitor-NC by Liposome transfection method.The cell proliferation ability was measured by CCK-8 at 48 h of culture,and the cell migra-tion ability was measured by Transwell migration experiment.At 24 h of culture,the ADAMTS1 protein was detected by Western blotting.HCASMC in the logarithmic growth phase were divided into groups A,B,C and D.Cells in the group A were transfected with mutant ADAMTS1 luciferase reporter gene plasmid(ADAMTS1-MUT)and miR-181a-5p mimics;cells in the group B were transfected with ADAMTS1-MUT and mimics-NC;cells in the group C were transfected with wild-type ADAMTS1 luciferase reporter gene plasmid(ADAMTS1-WT)and miR-181a-5p mimics;cells in the group D were transfected with ADAMTS1-WT and mimics-NC,respectively.The relative luciferase activity of cells in each group was de-tected by dual luciferase reporter gene assay at 24 h after culture.Results Compared with the control group,the relative expression of serum miR-181a-5p was lower in the ACS group(P<0.01);compared with UAP patients,the relative ex-pression of serum miR-181a-5p in AMI patients was lower(P<0.01).Compared with group Ⅱ,the proliferation and migra-tion abilities of cells decreased at 48 h after culture,and the relative expression of ADAMTS1 protein in cells of the group Ⅰ was lower at 24 h after culture(all P<0.01);compared with the group Ⅳ,the proliferation and migration abilities of cells were higher at 48 h after culture,and the relative expression of ADAMTS1 protein in cells of the group Ⅲ was higher at 24 h after culture(all P<0.01).The luciferase activity of cells in the groups A,B,C and D was 1.03±0.03,1.01±0.04,0.45±0.06,and 1.02±0.05,respectively;compared with the group B,the luciferase activity of cells in the group A was significantly lower(P<0.05).Conclusions The expression of miR-181a-5p is low in the serum of ACS patients.MiR-181a-5p can inhibit the proliferation and migration of HCASMC,and the mechanism may be that miR-181a-5p promotes the expression of ADAMTS1 protein in cells.
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