MicroRNA cloning and sequencing in osteosarcoma cell lines: differential role of miR-93

Cell Oncol (Dordr). 2012 Feb;35(1):29-41. doi: 10.1007/s13402-011-0059-z. Epub 2011 Sep 30.

Abstract

Background: Studies show that abnormalities in non-coding genes can contribute to carcinogenesis; microRNA levels may modulate cancer growth and metastatic diffusion.

Method: 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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics
  • Base Sequence
  • Cell Line, Tumor
  • Cell Proliferation
  • Clone Cells
  • Cloning, Molecular
  • E2F1 Transcription Factor / metabolism
  • Gene Expression Regulation, Neoplastic
  • Gene Library
  • Genetic Testing
  • Humans
  • Kinetics
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Molecular Sequence Data
  • Neoplasm Invasiveness
  • Osteosarcoma / genetics*
  • Osteosarcoma / pathology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Sequence Analysis, RNA*
  • Transendothelial and Transepithelial Migration / genetics
  • Transfection
  • Wound Healing

Substances

  • E2F1 Transcription Factor
  • E2F1 protein, human
  • MIRN93 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • RNA, Small Interfering