Department of Medical Examination & Health Management, First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, China.
Guangxi Key Laboratory of Precision Medicine in Cardio-cerebrovascular Diseases Control and Prevention & Guangxi Clinical Research Center for Cardio-cerebrovascular Diseases, Nanning, Guangxi, China.
Biomed Res Int. 2020 Aug 26;2020:9372961. doi: 10.1155/2020/9372961. eCollection 2020.
To explore the possible role of miR-499a-3p in the function of primary human umbilical vein endothelial cells (HUVECs) and the expression of ADAM10 in primary HUVEC.
miR-499a-3p was first transfected into primary HUVECs via lentivirus vector. The viability, proliferation, and migration of stable transfected primary HUVEC were then determined by flow cytometry, CCK8 assays, scratch tests, and Transwell tests. The transcription of miR-499a-3p and ADAM10 was examined by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of ADAM10 was examined by Western blot (WB).
After transfection, miR-499a-3p transcription was significantly increased ( < 0.01), compared to the blank and nonspecific control (NC) groups, while both ADAM10 transcription and expression were significantly decreased ( < 0.05). In contrast, in the inhibitors group, miR-499a-3p transcription was significantly reduced ( < 0.05) whereas both ADAM10 transcription and expression were significantly increased ( < 0.05). The viability, proliferation, and migration of primary HUVECs were significantly impaired ( < 0.05) by the transfection of miR-499a-3p but enhanced by miR-499a-3p inhibitors ( < 0.05).
Upregulation of miR-499a-3p transcription will inhibit the expression of ADAM10 in HUVECs; cell migration and proliferation, however, promote apoptosis. And reverse effects were established by downregulation of miR-499a-3p transcription. All these effects may be achieved by regulating the transcription and expression of ADAM10. These results combined suggested that miR-499a-3p may affect the proliferation, migration, and apoptosis of endothelial cells and regulate AS by regulating ADAM10. miR-499a-3p may become a candidate biomarker for the diagnosis of unstable angina pectoris (UA).
探讨微小 RNA-499a-3p（miR-499a-3p）在原代人脐静脉内皮细胞（HUVEC）功能中的可能作用及其在原代 HUVEC 中 ADAM10 的表达。
通过慢病毒载体将 miR-499a-3p 转染到原代 HUVEC 中。然后通过流式细胞术、CCK8 测定、划痕试验和 Transwell 试验测定稳定转染的原代 HUVEC 的活力、增殖和迁移。通过逆转录-聚合酶链反应（RT-PCR）检测 miR-499a-3p 和 ADAM10 的转录，通过 Western blot（WB）检测 ADAM10 的表达。
转染后，miR-499a-3p 转录明显增加（<0.01），与空白组和非特异性对照（NC）组相比，而 ADAM10 转录和表达均明显降低（<0.05）。相反，在抑制剂组中，miR-499a-3p 转录明显降低（<0.05），而 ADAM10 转录和表达均明显增加（<0.05）。miR-499a-3p 的转染明显损害了原代 HUVEC 的活力、增殖和迁移（<0.05），但 miR-499a-3p 抑制剂则增强了这些作用（<0.05）。
miR-499a-3p 转录上调会抑制 HUVEC 中 ADAM10 的表达；然而，细胞迁移和增殖促进细胞凋亡。下调 miR-499a-3p 转录则产生相反的效果。所有这些作用可能通过调节 ADAM10 的转录和表达来实现。这些结果表明，miR-499a-3p 可能通过调节 ADAM10 影响内皮细胞的增殖、迁移和凋亡，并通过调节 ADAM10 调节动脉粥样硬化。miR-499a-3p 可能成为不稳定型心绞痛（UA）诊断的候选生物标志物。
