环状 RNA PCNX 通过调控 miR-1278/DNMT1 轴促进人主动脉血管平滑肌细胞中 PDGF-BB 诱导的增殖和迁移。
CircPCNX Promotes PDGF-BB-Induced Proliferation and Migration of Human Aortic Vascular Smooth Muscle Cells Through Regulating miR-1278/DNMT1 Axis.
机构信息
Department of Laboratory Medicine, The First Hospital of Qiqihar/Affiliated Qiqihar Hospital, Southern Medical University, Qiqihar, 161005, Heilongjiang, China.
Department of Traditional Chinese Geriatric Medicine, The First Hospital of Qiqihar/Affiliated Qiqihar Hospital, Southern Medical University, Qiqihar, 161005, Heilongjiang, China.
出版信息
Cardiovasc Drugs Ther. 2023 Oct;37(5):877-889. doi: 10.1007/s10557-022-07342-y. Epub 2022 Jun 7.
BACKGROUND
Human aortic vascular smooth muscle cells (HA-VSMCs) play vital roles in the pathogenesis of vascular diseases. Circular RNAs (circRNAs) have been reported to regulate the biological functions of HA-VSMCs. In this study, the functions of circRNA pecanex homolog (circPCNX) in platelet-derived growth factor-BB (PDGF-BB)-induced HA-VSMCs were investigated.
METHODS
Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to determine the expression of circPCNX, DNA methyltransferase 1 (DNMT1), and microRNA-1278 (miR-1278). 5'-Ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry analysis, wound healing assay, and transwell assay were used to examine cell proliferation, cell cycle, and migration. Western blot assay was utilized to measure protein levels. RNA immunoprecipitation (RIP) assay, RNA pull down assay, and dual-luciferase reporter assay were adopted to analyze the relationships among circPCNX, miR-1278, and DNMT1.
RESULTS
CircPCNX was upregulated in PDGF-BB-treated HA-VSMCs in a dose- or time-dependent manner. CircPCNX knockdown alleviated PDGF-BB-induced cell proliferation, cell cycle progression, and migration in HA-VSMCs. CircPCNX knockdown could reverse PDGF-BB-induced HA-VSMC progression by regulating DNMT1. Moreover, circPCNX was identified to regulate DNMT1 expression by sponging miR-1278. Inhibition of miR-1278 reversed circPCNX knockdown-mediated effects on cell proliferation and migration in PDGF-BB-induced HA-VSMCs. MiR-1278 overexpression suppressed PDGF-BB-stimulated HA-VSMC proliferation and migration by targeting DNMT1.
CONCLUSION
CircPCNX promoted PDGF-BB-induced HA-VSMC proliferation and migration by elevating DNMT1 expression through sponging miR-1278.
背景
人主动脉血管平滑肌细胞（HA-VSMCs）在血管疾病的发病机制中起着至关重要的作用。环状 RNA（circRNA）已被报道可调节 HA-VSMCs 的生物学功能。在这项研究中，研究了 pecnex 同源物（circPCNX）在血小板衍生生长因子-BB（PDGF-BB）诱导的 HA-VSMCs 中的作用。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测 circPCNX、DNA 甲基转移酶 1（DNMT1）和 microRNA-1278（miR-1278）的表达。采用 5'-乙炔基-2'-脱氧尿苷（EdU）测定法、流式细胞术分析、划痕愈合试验和 Transwell 试验检测细胞增殖、细胞周期和迁移。采用 Western blot 检测蛋白水平。采用 RNA 免疫沉淀（RIP）试验、RNA 下拉试验和双荧光素酶报告基因试验分析 circPCNX、miR-1278 和 DNMT1 之间的关系。
结果
circPCNX 在 PDGF-BB 处理的 HA-VSMCs 中呈剂量或时间依赖性上调。circPCNX 敲低可减轻 PDGF-BB 诱导的 HA-VSMCs 增殖、细胞周期进程和迁移。circPCNX 敲低可通过调节 DNMT1 逆转 PDGF-BB 诱导的 HA-VSMC 进展。此外，circPCNX 通过海绵 miR-1278 来调节 DNMT1 的表达。抑制 miR-1278 逆转了 PDGF-BB 诱导的 circPCNX 敲低对 HA-VSMCs 增殖和迁移的影响。miR-1278 过表达通过靶向 DNMT1 抑制 PDGF-BB 刺激的 HA-VSMC 增殖和迁移。
结论
circPCNX 通过海绵 miR-1278 升高 DNMT1 表达促进 PDGF-BB 诱导的 HA-VSMC 增殖和迁移。
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