title: MicroRNA-1298 is regulated by DNA methylation and affects vascular smooth muscle cell function by targeting connexin 43.
abstract: OBJECTIVE
  Growing evidence links microRNA to the process of peripheral vascular disease. Recently, we have found that microRNA-1298(miR-1298) is one of the most significantly down-regulated microRNAs in human arteries with arteriosclerosis obliterans (ASO) of the lower extremities. However, little is known regarding its role in the process of ASO. The present study aimed to investigate the expression, regulatory mechanisms, and functions of miR-1298 in the process of ASO.
  
  
  RESULTS
  Using quantitative reverse-transcription PCR and in situ hybridization assays, miR-1298 was observed predominantly expressed in the vascular smooth muscle cells (VSMCs) and was significantly down-regulated in ASO compared with normal arteries. Pyrosequencing analysis revealed that the miR-1298 DNA upstream of CpG sites were hypermethylated in ASO compared with normal arteries. Next, the luciferase reporter assay revealed that miR-1298 down-regulation is related with upstream DNA CpG site hypermethylation. Introducing a miR-1298 mimic into cultured VSMCs significantly attenuated cell proliferation and migration. Connexin 43 (Cx43) was validated to be a functional target of miR-1298 that was involved in the miR-1298-mediated cellular effects. Finally, lentivirus-mediated delivery of miR-1298 and its target Cx43 into a rat carotid balloon injury model indicated that re-overexpression of miR-1298 significantly decreased neointimal formation by targeting connexin 43.
  
  
  CONCLUSIONS
  Our data demonstrate a specific role of the upstream DNA methylation/miR-1298/Cx43 pathway in regulating VSMC function and suggest that modulation of miR-1298 levels may offer a novel therapeutic approach for ASO.
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China.
authors.forename: Mian
authors.initials: M
authors.lastname: Wang
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China.
authors.forename: Henghui
authors.initials: H
authors.lastname: Yin
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Yao
authors.affiliation: Department of Pathology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: He
authors.affiliation: Division of Vascular Surgery, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, China.
authors.forename: Leping
authors.initials: L
authors.lastname: Yin
authors.affiliation: Department of Pharmacology, Rush University Medical Center, Chicago, USA.
authors.forename: Chunxiang
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Laboratory of General Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Li
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China 13922231628@163.com shenmingwang@hotmail.com.
authors.forename: Guangqi
authors.initials: G
authors.lastname: Chang
authors.affiliation: Division of Vascular Surgery, The Guangdong Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The Vascular Surgical Disease Research Center of Guangdong Province, First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shan Er Road, Guangzhou, Guangdong 510080, China 13922231628@163.com shenmingwang@hotmail.com.
authors.forename: Shenming
authors.initials: S
authors.lastname: Wang
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism*; D019175:DNA Methylation; D015536:Down-Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D058426:Neointima / Q000378:metabolism
chemical_list: D018031:Connexin 43; C500216:GJA1 protein, human; C000609803:MIRN1298 microRNA, human; D035683:MicroRNAs
keywords: Arteriosclerosis obliterans; Connexin 43; Methylation; Smooth muscle cell; microRNA