Screening of miRNAs Associated with Hcy Promoting Proliferation and Migration of Vascular Smooth Muscle Cells
Objective To determine the effects of homocysteine(Hcy)on proliferation and migration of human vascular smooth muscle cell(VSMC)and analyze the differences related to the proliferation and migration of VSMC under the intervention of Hcy.Methods VSMC was cultured in v itro and divided into Control group and 100 μmol·L-1 Hcy intervention group.The proliferative activity of VSMC was measured by CCK-8 method.The migration of VSMC at 0 h and 48 h was analyzed by scratch test.The differentiated expression of miRNAs in VSMC in each group was analyzed by high-throughput sequencing,target gene prediction was performed us-ing miRanda and TargetScan software,and the function of target genes was analyzed by BLAST software com-pared with GO and KEGG databases to obtain the target gene information related to VSMC proliferation and mi-gration.The up-regulated miRNAs were verified by RT-q PCR.Results Hcy intervention could significantly increase the proliferative activity of VSMC(P<0.01),and the scratch area of VSMC was significantly reduced(P<0.01),suggesting that 100 μmol·L-1 Hcy intervention could promote the proliferation and migration of VSMC,which was consistent with the results of previous studies.In terms of mechanism,a total of 576 miRNAs were detected in both groups of cells,of which 404 were known and 172 were newly predicted.Compared with the Control group,there were 88 differentially expressed miRNAs in the Hcy group,of which 20 were up-regu-lated and 68 were down-regulated.Among them,there were 20 miRNAs related to VSMC proliferation and mi-gration,including miR NA39,miR NA206,and miRNA212-5p whose expression was up-regulated,and 17 miR NA35 whose expression was down-regulated.The results of RT-qPCR showed that the expressions of miR-NA212-5p was significantly increased in the Hcy group(P<0.05).Conclusion There are differentially ex-pressed miRNAs in the proliferation and migration of VSMC induced by Hcy.miR NA212-5p may be involved in the effect of Hcy on VSMC.
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