title: Inhibitory roles of miR-320 in osteosarcoma via regulating E2F1.
abstract: OBJECTIVE
  Osteosarcoma (OsC) is the most common primary bone malignant tumor with lower incidence and high degree of malignancy, but the exact mechanism remains unknown. More evidence demonstrated microRNAs (miRNAs) could contribute to tumor progression. In this study, we investigated the expression and functions of miR-320 in OsC cells.
  
  
  METHODS
  miR-320 expression levels in several human OsC cell lines and human normal osteoblastic cell line were tested by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). U2OS cells were transfected with miR-320 mimics or negative control oligos. MTT assay and cell flow cytometry assay by PI staining were performed to access the cell growth rate. Bioinformatic prediction and luciferase assays were used to identify the predicted target E2F1. qRT-PCR and Western blot were performed to access the molecular alteration of E2F1.
  
  
  RESULTS
  miR-320 was decreased in human OsC cell lines. Heterogeneous expression of miR-320 inhibited cell proliferation and induced cell cycle arrest. Besides, we proved that miR-320 could directly regulate the expression of E2F1 in U2OS cells.
  
  
  CONCLUSIONS
  These data suggested that miR-320 regulates the proliferation and cell cycle by targeting E2F1 in human OsC progression.
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Haojie
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Minghui
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Shutao
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Dengfeng
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Henan University Huaihe Hospital, Kaifeng 475000, Henan, P.R. China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D001665:Binding Sites; D059447:Cell Cycle Checkpoints / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018450:Disease Progression; D050687:E2F1 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D012516:Osteosarcoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference
chemical_list: D020413:3' Untranslated Regions; D050687:E2F1 Transcription Factor; C572916:MIRN320 microRNA, human; D035683:MicroRNAs