Chen Hua, Liu Xi, Wu Yun, Wu Xiayin, Wen Xiaoli, Lu Yanan, Zhao Xingsheng
Department of Cardiology, Inner Mongolia People's Hospital, Hohhot, Inner Mongolia 010000, P.R. China.
Exp Ther Med. 2021 Jun;21(6):661. doi: 10.3892/etm.2021.10093. Epub 2021 Apr 21.
Atherosclerosis causes stroke and coronary heart disease and is associated with a high mortality rate worldwide. However, the pathogenesis of atherosclerosis remains unclear. Endothelial cell apoptosis is one of the early changes observed in atherosclerosis. Previous studies have found that microRNA (miR)-616-3p may be involved in the development of atherosclerosis, but the specific mechanism is not clear. The present study aimed to investigate whether miR-616-3p is involved in endothelial cell apoptosis and its underlying mechanism. The present study demonstrated that compared with normal HUVECs, HUVECs treated with oxidized low-density lipoprotein expressed higher miR-616-3p and lower X-linked inhibitor of apoptosis protein (XIAP) levels. In the present study, HUVECs were transfected with miR-616-3p mimic and Cell Counting Kit-8 (CCK-8), flow cytometry and TUNEL staining assays demonstrated that compared with miR-616-3p mimic control, the miR-616-3p mimic promoted HUVEC apoptosis. In addition, using StarBase 3.0 for bioinformatics analysis it was predicted that miR-616-3p may bind to the 3'untranslated region (UTR) of XIAP mRNA. The present study performed the CCK-8, flow cytometry, TUNEL staining and dual-luciferase reporter assays and demonstrated that miR-616-3p binds to the 3'UTR of the XIAP mRNA and inhibits its expression and that this further promotes apoptosis in HUVECs. In addition, western blotting demonstrated that compared with miR-616-3p mimic control, the miR-616-3p mimic increases the level of cleaved caspase-3 in HUVECs. In summary, the present study demonstrated that miR-616-3p can directly inhibit the expression of XIAP mRNA by targeting its 3'UTR which promoted apoptosis in HUVECs. miR-616-3p and XIAP may be used as therapeutic targets of atherosclerosis in the future.
动脉粥样硬化可导致中风和冠心病，在全球范围内与高死亡率相关。然而，动脉粥样硬化的发病机制仍不清楚。内皮细胞凋亡是在动脉粥样硬化中观察到的早期变化之一。先前的研究发现，微小RNA（miR）-616-3p可能参与动脉粥样硬化的发展，但其具体机制尚不清楚。本研究旨在探讨miR-616-3p是否参与内皮细胞凋亡及其潜在机制。本研究表明，与正常的人脐静脉内皮细胞（HUVECs）相比，用氧化型低密度脂蛋白处理的HUVECs表达更高的miR-616-3p和更低的X连锁凋亡抑制蛋白（XIAP）水平。在本研究中，用miR-616-3p模拟物转染HUVECs，细胞计数试剂盒-8（CCK-8）、流式细胞术和TUNEL染色分析表明，与miR-616-3p模拟物对照相比，miR-616-3p模拟物促进了HUVEC凋亡。此外，使用StarBase 3.0进行生物信息学分析预测，miR-616-3p可能与XIAP mRNA的3'非翻译区（UTR）结合。本研究进行了CCK-8、流式细胞术、TUNEL染色和双荧光素酶报告基因分析，并表明miR-616-3p与XIAP mRNA的3'UTR结合并抑制其表达，进而进一步促进HUVEC凋亡。此外，蛋白质印迹法表明，与miR-616-3p模拟物对照相比，miR-616-3p模拟物增加了HUVECs中裂解的半胱天冬酶-3的水平。总之，本研究表明，miR-616-3p可通过靶向XIAP mRNA的3'UTR直接抑制其表达，从而促进HUVEC凋亡。miR-616-3p和XIAP未来可能用作动脉粥样硬化的治疗靶点。
