title: miR-124 suppresses the migration and invasion of glioma cells in vitro via Capn4.
abstract: miR-124 and Capn4 are aberrantly expressed in glioblastoma multiforme (GBM) tissues. In the present study, we investigated miR-124 and Capn4 expression in GBM tissue specimens. The role of miR-124 and Capn4 in the migration and invasion of glioma cells in vitro was also examined. miR-124 and Capn4 expression in 20 GBM and 6 control brain specimens was examined using RT-qPCR and immuno-blotting. Data from The Cancer Genome Atlas were retrieved. Candidate mRNA target sites of miR-124 were predicted using TargetScan/microRNA and binding was examined using dual luciferase reporter assays. The U87 and U251 cells were transfected with scrambled microRNA, miR-124 mimics and/or pLenti-Capn4 prior to wound‑healing and Transwell invasion assays. Proteins involved in the epithelial-mesenchymal transition were examined using immunoblotting. The results showed that miR-124 was significantly downregulated in GBM tissues. Immunoblotting showed a marked upregulation of Capn4 expression in GBM tissues. The Spearman's correlation analysis revealed a negative association between miR-124 expression and Capn4 protein levels. TargetScan/microRNA predicted the miR-124 binding site in the nucleotide 440-446 region within the Capn4 3'-UTR, which was confirmed by luciferase assays. Wound‑healing and Transwell invasion assays demonstrated that Capn4 downregulation or miR-124 mimics suppressed the migration and invasion of glioma cells. Capn4 downregulation or miR-124 mimics reduced the level of phospho-FAK and MMP2, vimentin and N-cadherin in U87 cells. In conclusion, miR-124 was found to suppress the migration and invasion of glioma cells in vitro via Capn4.
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Jia-Jun
authors.initials: JJ
authors.lastname: Cai
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Zeng-Xin
authors.initials: ZX
authors.lastname: Qi
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Ling-Chao
authors.initials: LC
authors.lastname: Chen
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Yao
authors.affiliation: Department of Geriatrics, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gong
authors.affiliation: Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Mao
mesh_terms: D020413:3' Untranslated Regions; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D058750:Epithelial-Mesenchymal Transition; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D066298:In Vitro Techniques; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009361:Neoplasm Invasiveness
chemical_list: D020413:3' Untranslated Regions; C509207:MIRN124 microRNA, human; D035683:MicroRNAs; D002154:Calpain; C527669:CAPNS1 protein, human