Clinical Biochemistry Department, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran; Student Research Committee, Iran University of Medical Sciences, Tehran, Iran.
Neuroscience Research Center, Vice-Chancellor for Research and Technology, Iran University of Medical Sciences, Tehran, Iran; Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.
Microvasc Res. 2021 Jul;136:104172. doi: 10.1016/j.mvr.2021.104172. Epub 2021 Apr 21.
VSMC proliferation and migration pathways play important roles in plaque formation in the vessel stenosis and re-stenosis processes. The microRNAs affect the expression of many genes that regulate these cellular processes. The aim of this study was to investigate the effects of miR-181b, miR-204, and miR-599 on the gene and protein expression levels of hematopoietic cell kinase (HCK) in VSMCs.
miR-181b, miR-204 were predicted for the suppression of HCK in the chemokine signaling pathway using bioinformatics tools. Then, the VSMCs were transfected by PEI-containing microRNAs. The HCK gene and protein expression levels were evaluated using RT-qPCR and Western blotting techniques, respectively. Moreover, the cellular proliferation and migration were evaluated by MTT and scratch assay methods.
The miR-181b and miR-204 decreased significantly the HCK gene and (total and phosphorylated) protein expression levels. Also, the miR-599 did not show any significant effects on the HCK gene and protein levels. The data also showed that miR-181b, miR-204, and miR-599 prevent significantly the proliferation and migration of VSMCs.
The downregulation of HCK by miR-181b and miR-204 suppressed the VSMC proliferation and migration.
血管平滑肌细胞（VSMC）增殖和迁移途径在血管狭窄和再狭窄过程中的斑块形成中起着重要作用。微小 RNA（miRNA）影响调节这些细胞过程的许多基因的表达。本研究旨在探讨 miR-181b、miR-204 和 miR-599 对 VSMCs 中造血细胞激酶（HCK）基因和蛋白表达水平的影响。
使用生物信息学工具预测 miR-181b 和 miR-204 在趋化因子信号通路中对 HCK 的抑制作用。然后，通过含有 PEI 的 microRNAs 转染 VSMCs。分别采用 RT-qPCR 和 Western blot 技术评估 HCK 基因和蛋白表达水平。此外，通过 MTT 和划痕实验方法评估细胞增殖和迁移。
miR-181b 和 miR-204 显著降低了 HCK 基因和（总蛋白和磷酸化蛋白）的表达水平。此外，miR-599 对 HCK 基因和蛋白水平没有显示出任何显著影响。数据还表明，miR-181b、miR-204 和 miR-599 可显著抑制 VSMC 的增殖和迁移。
miR-181b 和 miR-204 通过下调 HCK 抑制了 VSMC 的增殖和迁移。
