title: Mitomycin C induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-200b and its targeting of RhoE.
abstract: Mitomycin C (MMC) is known to reduce epidural fibrosis, but the underlying mechanisms have not yet been elucidated. Aberrant miR-200b expressions have been reported in multiple types of fibrotic tissues from many diseases. The aim of this study was to clarify the mechanism by which MMC induces fibroblasts apoptosis and reduces epidural fibrosis. The expression of miR-200b in human fibroblasts was determined after MMC treatment, and the targeted association between miR-200b and RhoE was determined using the luciferase activity assay. The effects of MMC and miR-200b on human fibroblasts apoptosis were evaluated using flow cytometry and western blot analysis. The effects of MMC and miR-200b on epidural fibrosis were evaluated using the Rydell classification, hydroxyproline content, apoptotic cell count and histological analysis. The study revealed that MMC could significantly downregulate miR-200b expression and induce human fibroblasts apoptosis. The direct downregulation of miR-200b could induce human fibroblasts apoptosis. Furthermore, we identified the binding sequence for miR-200b within the 3' untranslated region of RhoE. RhoE was confirmed to be a direct target of miR-200b, and RhoE itself acted as a promoter of fibroblasts apoptosis. The inhibition of miR-200b increased fibroblasts apoptosis and reduced epidural fibrosis in rats, which was in accordance with the effect of MMC. This study suggests that MMC induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-200b expression and its targeting of RhoE.
authors.affiliation: Department of Orthopedics, Clinical Medical College of Yangzhou University, No. 98 Nantong West Road, Yangzhou 225001, China; Department of Microbiology, Nanjing Medical University, No. 140 Hanzhong Road, Nanjing 210029, China; Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou Road, Nanjing 210029, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Physiology, Nanjing Medical University, No. 140 Hanzhong Road, Nanjing 210029, China.
authors.forename: Yingbin
authors.initials: Y
authors.lastname: Ge
authors.affiliation: Department of Physiology, Nanjing Medical University, No. 140 Hanzhong Road, Nanjing 210029, China.
authors.forename: Yuxuan
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Department of Orthopedics, Clinical Medical College of Yangzhou University, No. 98 Nantong West Road, Yangzhou 225001, China.
authors.forename: Lianqi
authors.initials: L
authors.lastname: Yan
authors.affiliation: Department of Orthopedics, Clinical Medical College of Yangzhou University, No. 98 Nantong West Road, Yangzhou 225001, China; Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou Road, Nanjing 210029, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Cai
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou Road, Nanjing 210029, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Shi
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou Road, Nanjing 210029, China. Electronic address: xiaojiancao@gmail.com.
authors.forename: Xiaojian
authors.initials: X
authors.lastname: Cao
authors.affiliation: Department of Microbiology, Nanjing Medical University, No. 140 Hanzhong Road, Nanjing 210029, China. Electronic address: clu@njmu.edu.cn.
authors.forename: Chun
authors.initials: C
authors.lastname: Lu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D002921:Cicatrix / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D015536:Down-Regulation; D004824:Epidural Space; D055111:Failed Back Surgery Syndrome / Q000209:etiology / Q000473:pathology / Q000517:prevention & control; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology; D005355:Fibrosis; D057809:HEK293 Cells; D006801:Humans; D007796:Laminectomy / Q000009:adverse effects; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D016685:Mitomycin / Q000494:pharmacology*; D061251:Primary Cell Culture; D017207:Rats, Sprague-Dawley; D020741:rho GTP-Binding Proteins / Q000037:antagonists & inhibitors*
chemical_list: C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D016685:Mitomycin; C508705:RND3 protein, human; D020741:rho GTP-Binding Proteins
keywords: Epidural fibrosis; Fibroblasts apoptosis; Mitomycin C; RhoE; miR-200b