miR-205 expression promotes cell proliferation and migration of human cervical cancer cells

PLoS One. 2012;7(10):e46990. doi: 10.1371/journal.pone.0046990. Epub 2012 Oct 3.

Abstract

MicroRNAs (miRNAs) are short non-coding RNA regulators that control gene expression mainly through post-transcriptional silencing. We previously identified miR-205 in a signature for human cervical cancer using a deep sequencing approach. In this study, we confirmed that miR-205 expression was frequently higher in human cervical cancer than their matched normal tissue samples. Functionally, we demonstrate that miR-205 promotes cell proliferation and migration in human cervical cancer cells. To further understand the biological roles of miR-205, we performed in vivo crosslinking and Argonaute 2 immunoprecipitation of miRNA ribonucleoprotein complexes followed by microarray analysis (CLIP-Chip) to identify its potential mRNA targets. Applying CLIP-Chip on gain- and loss-of-function experiments, we identified a set of transcripts as potential targets of miR-205. Several targets are functionally involved in cellular proliferation and migration. Two of them, CYR61 and CTGF, were further validated by Western blot analysis and quantification of mRNA enrichment in the Ago2 immunoprecipitates using qRT-PCR. Furthermore, both CYR61 and CTGF were downregulated in cervical cancer tissues. In summary, our findings reveal novel functional roles and targets of miR-205 in human cervical cancer, which may provide new insights about its role in cervical carcinogenesis and its potential value for clinical diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Movement / genetics*
  • Cell Proliferation
  • Connective Tissue Growth Factor / genetics
  • Cysteine-Rich Protein 61 / genetics
  • Female
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism*
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / metabolism
  • Uterine Cervical Neoplasms / pathology*

Substances

  • CCN2 protein, human
  • Cysteine-Rich Protein 61
  • MIRN205 microRNA, human
  • MicroRNAs
  • Connective Tissue Growth Factor

Grant support

This work was supported by the Swedish Research Council (523-2009-3517, 521-2010-3518); the Åke Olsson’s Foundation for Haematological Research; the Swedish Cancer Foundation; the Åke Wiberg’s Foundation; King Gustaf V Jubilee Fund; Cancer Society in Stockholm; the Axel and Signe Lagerman’s Donation Foundation; Karolinska Institutet and Stockholm County Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.