目的:血管生成在冠状动脉侧支循环(CCC)中起关键作用,慢性冠状动脉完全闭塞期间新血管的代偿形成。本研究旨在确定浆细胞瘤变异型易位1(PVT1),参与肿瘤血管生成的长链非编码(lnc)RNA,在慢性冠脉缺血期间对血管生成有调节作用。
方法:冠心病患者,狭窄≥90%,根据RentropCohen分类,将其分为“好”和“差”CCC组。从所有患者获得RNA样本,以及来自缺氧和葡萄糖剥夺(OGD)HUVECs。用RT-qPCR或Westernblot检测PVT1、miR-15b-5p和AKT3水平,而HUVEC迁移和血管生成被检测到,分别,伤口愈合和管形成测定。荧光素酶报告基因测定证实了PVT1-miR-15b-5p的直接结合。
结果:在“良好的CCC”患者血浆中发现PVT1升高,随着在OGDHUVEC中高表达;PVT1敲低降低HUVEC迁移,管形成,和促血管生成因子的表达。相反,OGDHUVECs下调miR-15b-5p,和miR-15b-5p过表达显著降低了它们的血管生成能力。这些PVT1敲低或miR-15b-5p过表达相关的血管生成作用的减少被AKT3过表达逆转。在体内,注射ADV-sh-PVT1的缺血小鼠后肢和梗死心肌的新生血管形成和功能均减少,通过同时注射antagomiR-15b-5p来改善。
结论:循环PVT1可以作为一种有用的生物标志物来区分好的和差的CCC,因为它参与通过miR-15b-5p-AKT3轴协调血管生成;因此它有可能作为治疗缺血性疾病的靶标。
OBJECTIVE: Angiogenesis plays a key role in coronary collateral circulation (CCC), the compensatory formation of new blood vessels during chronic total coronary occlusion. This study aimed to determine whether plasmacytoma variant translocation 1 (PVT1), a long non-coding (lnc) RNA involved in tumor angiogenesis, plays a role in regulating angiogenesis during chronic coronary ischemia.
METHODS: Patients with coronary artery disease, and ≥ 90% stenosis, were examined and divided into \"Good\" and \"Poor\" CCC groups based on Rentrop Cohen classification. RNA samples were obtained from all patients, as well as from oxygen and glucose-deprived (OGD) HUVECs. PVT1, miR-15b-5p and AKT3 levels were measured with RT-qPCR or Western blot, while HUVEC migration and angiogenesis were detected by, respectively, wound-healing and tube formation assays. Luciferase reporter assay confirmed direct PVT1-miR-15b-5p binding.
RESULTS: Increased PVT1 was found in \"Good CCC\" patient plasma, along with being highly expressed among OGD HUVECs; PVT1 knockdown reduced HUVEC migration, tube formation, and pro-angiogenic factor expression. Conversely, OGD HUVECs had downregulated miR-15b-5p, and miR-15b-5p overexpression significantly depressed their angiogenic capabilities. These PVT1 knockdown- or miR-15b-5p overexpression-associated reductions in angiogenic effects were reversed by AKT3 overexpression. In vivo, neovascularization and functioning in both ischemic mice hind-limbs and infarcted myocardium injected with ADV-sh-PVT1 were reduced, which were ameliorated by concurrent antagomiR-15b-5p injections.
CONCLUSIONS: Circulating PVT1 may serve as a useful biomarker to distinguish between good versus poor CCC, as it is involved in orchestrating angiogenesis via the miR-15b-5p-AKT3 axis; it thus has potential as a target for treating ischemic disease.