title: MicroRNA-145 regulates platelet-derived growth factor-induced human aortic vascular smooth muscle cell proliferation and migration by targeting CD40.
abstract: The objective of this study is to investigate the expression of microRNA (miR)-145 in human aortic vascular smooth muscle cells (VSMCs) and the effect of miR-145 in the biological behavior and expression of CD40 in VSMCs. Cells were treated with either miR-145 or miR-145 inhibitor. Cell proliferation was analyzed by a colony formation assay and a methyl thiazolyl tetrazolium assay. Cell migration and invasion were assessed using a transwell assay, an invasion assay, and a wound healing assay. A luciferase reporter assay was used to detect the interaction between miR-145 and CD40. Expression of α-SMA, calponin, osteopontin (OPN), epiregulin, activator protein-1 (AP-1) and CD40 was measured using real-time RT-PCR for mRNA levels and Western blotting for protein levels. Overexpression of miR-145 significantly inhibited VSMC proliferation, invasion and migration. Furthermore, OPN, epiregulin, AP-1 and CD40 expression at the mRNA and protein levels was down-regulated by overexpression of miR-145. However, α-SMA and calponin expression at the mRNA and protein levels was up-regulated by overexpression of miR-145. In addition, the luciferase reporter assay showed that CD40 may be a direct target gene of miR-145 in VSMC initiation and development. Moreover, these data demonstrate that the up-regulation of CD40 is critical for miR-145-mediated inhibitory effects on platelet-derived growth factor-induced cell proliferation and migration in human VSMCs. In summary, CD40, a direct target of miR-145, reverses the inhibitory effects of miR-145. These results suggest that the specific modulation of miR-145 in human VSMCs may be an attractive approach for the treatment of proliferative vascular diseases.
authors.affiliation: Centre for Cellular & Structural Biology, Sun Yat-Sen University Guangzhou City, Guangdong Province, P. R. China.
authors.forename: Yumei
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Hypertension, The First Affiliated Hospital of Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Jiangnan
authors.initials: J
authors.lastname: Huang
authors.affiliation: Department of Hypertension, The First Affiliated Hospital of Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: Department of Hypertension, The First Affiliated Hospital of Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Yuanli
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: Department of Hypertension, The First Affiliated Hospital of Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Mingjie
authors.initials: M
authors.lastname: Xia
authors.affiliation: College of Pharmacy, Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Pharmacy, Guangxi Medical University Nanning City, Guangxi Province, P. R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Jiao
keywords: CD40; microRNA-145; migration; platelet-derived growth factor; proliferation; vascular smooth muscle cells