title: MicroRNA cloning and sequencing in osteosarcoma cell lines: differential role of miR-93.
abstract: BACKGROUND
  Studies show that abnormalities in non-coding genes can contribute to carcinogenesis; microRNA levels may modulate cancer growth and metastatic diffusion.
  
  
  METHODS
  MicroRNA libraries were built and sequenced from two osteosarcoma cell lines (MG-63 and 143B), which differ in proliferation and transmigration. By cloning and transfection, miR-93, expressed in both cell lines, was then investigated for its involvement in osteosarcoma progression.
  
  
  RESULTS
  Six of the 19 miRNA identified were expressed in both cell lines with higher expression levels of miR-93 in 143B and in primary osteosarcoma cultures compared to normal osteoblasts. Interestingly, levels of miR-93 were significantly higher in metastases from osteosarcoma than in paired primary tumours. When 143B and MG-63 were transfected with miR-93, clones appeared to respond differently to microRNA overexpression. Ectopic expression of miR-93 more significantly increased cell proliferation and invasivity in 143B than in MG-63 clones. Furthermore, increased mRNA and protein levels of E2F1, one of the potential miR-93 targets, were seen in osteosarcoma cellular clones and its involvement in 143B cell proliferation was confirmed by E2F1 silencing.
  
  
  CONCLUSIONS
  Although further studies are needed to evaluate miRNA involvement in osteosarcoma progression, miR-93 overexpression seems to play an important role in osteosarcoma cell growth and invasion.
authors.affiliation: Center for Molecular and Translational Oncology, Department of Genetics, Microbiology, Antropology and Evolution, University of Parma, Parma, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Montanini
authors.forename: Lisa
authors.initials: L
authors.lastname: Lasagna
authors.forename: Valeria
authors.initials: V
authors.lastname: Barili
authors.forename: Søren Peter
authors.initials: SP
authors.lastname: Jonstrup
authors.forename: Alba
authors.initials: A
authors.lastname: Murgia
authors.forename: Laura
authors.initials: L
authors.lastname: Pazzaglia
authors.forename: Amalia
authors.initials: A
authors.lastname: Conti
authors.forename: Chiara
authors.initials: C
authors.lastname: Novello
authors.forename: Jørgen
authors.initials: J
authors.lastname: Kjems
authors.forename: Roberto
authors.initials: R
authors.lastname: Perris
authors.forename: Maria Serena
authors.initials: MS
authors.lastname: Benassi
mesh_terms: D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002999:Clone Cells; D003001:Cloning, Molecular; D050687:E2F1 Transcription Factor / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D005820:Genetic Testing; D006801:Humans; D007700:Kinetics; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D012516:Osteosarcoma / Q000235:genetics* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D017423:Sequence Analysis, RNA; D057705:Transendothelial and Transepithelial Migration / Q000235:genetics; D014162:Transfection; D014945:Wound Healing
chemical_list: D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; C543350:MIRN93 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering