CircHIPK3 Regulates Vascular Smooth Muscle Cell Calcification Via the miR-106a-5p/MFN2 Axis

• Published in: Journal of cardiovascular translational research 2022-12, Vol.15 (6), p.1315-1326
• Main Authors: Zhang, Wen-Bo, Qi, You-Fei, Xiao, Zhan-Xiang, Chen, Hao, Liu, Sa-Hua, Li, Zhen-Zhen, Zeng, Zhao-Fan, Wu, Hong-Fei
• Format: Article
• Language: English
• Subjects: Atherosclerosis - genetics; Biomedical Engineering and Bioengineering; Biomedicine; Calcium; Cardiology; Cell Differentiation; GTP Phosphohydrolases - genetics; Human Genetics; Humans; Medicine; Medicine & Public Health; MicroRNAs - genetics; MicroRNAs - metabolism; Mitochondrial Proteins - genetics; Muscle, Smooth, Vascular - metabolism; Original Article; Osteogenesis - genetics; RNA, Circular - genetics; Vascular Calcification - genetics
• ISSN: 1937-5387; 1937-5395
• DOI: 10.1007/s12265-022-10247-8

Atherosclerosis is the most common arterial disease and is closely related to vascular calcification. CircHIPK3 has been implicated in atherosclerosis development, but the possible downstream regulatory mechanisms remain unclear. The levels of circHIPK3, miR-106a and MFN2 in tissues and blood samples of patients with atherosclerosis were detected by RT-qPCR. The levels of circHIPK3, miR-106a and MFN2 were detected by RT-qPCR and the expression levels of MFN2, osteogenic and cartilage differentiation marker proteins were detected by western blot in vitro. ALP staining, Alizarin Red staining, and calcium content detection evaluated the degree of osteogenic differentiation of cells. Alcian blue staining detected the level of cell cartilage differentiation. Luciferase detected the targeting relationship between circHIPK3 and miR-106a-5p, as well as miR-106a-5p and MFN2. CircHIPK3 and MFN2 were low expressed and miR-106a-5p was highly expressed in tissues and blood samples of patients with atherosclerosis, as well as vascular smooth muscle cell (VSMC) with osteogenic and cartilage differentiation. Overexpression of circHIPK3 reduced the cell mineralization and calcium content. Overexpression of circHIPK3 inhibited osteogenic differentiation by decreasing ALP activity, RUNX2, and OPG expression, and increasing SM22α and SMA level. What’s more, overexpression of circHIPK3 decreased the chondrogenic differentiation by inhibiting the protein level of SOX9, aggrecan, and collagen II. CircHIPK3 targeted miR-106a-5p and miR-106a-5p targeted MFN2. MiR-106a-5p overexpression or MFN2 depletion repressed the effect of circHIPK3 overexpression on VSMC calcification. CircHIPK3 regulated osteogenic and cartilage differentiation of VSMC via miR-106a-5p/MFN2 axis, indicating a target for treating vascular calcification.