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siRNA Mediated Silencing of NIN1/RPN12 Binding Protein 1 Homolog Inhibits Proliferation and Growth of Breast Cancer Cells

  • Huang, Wei-Yi (Department of Oncology, Affiliated People's 1st Hospital, Shanghai Jiaotong University) ;
  • Chen, Dong-Hui (Department of Oncology, Affiliated People's 1st Hospital, Shanghai Jiaotong University) ;
  • Ning, Li (Department of Oncology, Affiliated People's 1st Hospital, Shanghai Jiaotong University) ;
  • Wang, Li-Wei (Department of Oncology, Affiliated People's 1st Hospital, Shanghai Jiaotong University)
  • Published : 2012.05.30

Abstract

The gene encoding the Nin one binding (NOB1) protein which plays an essential role in protein degradation has been investigated for possible tumor promoting functions. The present study was focused on NOB1 as a possible therapeutic target for breast cancer treatment. Lentivirus mediated NOB1 siRNA transfection was used to silence the NOB1 gene in two established breast cancer cell lines, MCF-7 and MDA-MB-231, successful transfection being confirmed by fluorescence imaging. NOB1 deletion caused significant decline in cell proliferation was observed in both cell lines as investigated by MTT assay. Furthermore the number and size of the colonies formed were also significantly reduced in the absence of NOB1. Moreover NOB1 gene knockdown arrested the cell cycle and inhibited cell cycle related protein expression. Collectively these results indicate that NOB1 plays an essential role in breast cancer cell proliferation and its gene expression could be a therapeutic target.

Keywords

References

  1. Adams J (2004). The proteasome: a suitable antineoplastic target. Nat Rev Cancer, 4, 349-60. https://doi.org/10.1038/nrc1361
  2. Fasanaro P, Capogrossi MC, Martelli F (2010). Regulation of the endothelial cell cycle by the ubiquitin-proteasome system. Cardiovasc Res, 85, 272-80. https://doi.org/10.1093/cvr/cvp244
  3. Ferrell K, Wilkinson CR, Dubiel W, Gordon C (2000). Regulatory subunit interactions of the 26S proteasome, a complex problem. Trends Biochem Sci, 25, 83-8. https://doi.org/10.1016/S0968-0004(99)01529-7
  4. Hanahan Da WRA (2000). The hallmarks of cancer. Cell, 100, 57-70. https://doi.org/10.1016/S0092-8674(00)81683-9
  5. Hershko Aa CA (1998). The ubiquitin system. Annual Review Biochem, 67, 425-79. https://doi.org/10.1146/annurev.biochem.67.1.425
  6. Lin Y, Peng S, Yu H, Teng H, Cui M (2012). RNAi-mediated downregulation of NOB1 suppresses the growth and colonyformation ability of human ovarian cancer cells. Med Oncol, 29, 311-7. https://doi.org/10.1007/s12032-010-9808-5
  7. Lu Z, Guo Q, Shi A, Xie F, Lu Q (2012). Down regulation of NIN/RPN12 binding protein inhibit the growth of human hepatocellular carcinoma cells. Mol Biol Rep, 39, 501-7. https://doi.org/10.1007/s11033-011-0764-8
  8. Mello CC, Conte D Jr. (2004). Revealing the world of RNA interference. Nature, 431, 338-42. https://doi.org/10.1038/nature02872
  9. Pertschy B, Schneider C, Gnadig M, et al (2009). RNA helicase prp43 and its co-factor Pfa1 promote 20 to 18S rRNA processing catalyzed by the endonuclease Nob1. J Biol Chem, 284, 35079-91. https://doi.org/10.1074/jbc.M109.040774
  10. Shirane M, Harumiya Y, Ishida N, et al (1999). Down-regulation of p27(Kip1) by two mechanisms, ubiquitin-mediated degradation and proteolytic processing. J Biol Chem, 274, 13886-93. https://doi.org/10.1074/jbc.274.20.13886
  11. Soneoka Y, Cannon PM, Ramsdale E, et al (1995). A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res, 23, 628-33. https://doi.org/10.1093/nar/23.4.628
  12. Storvold GL, Andersen TI, Perou CM, Frengen E (2006). siRNA: a potential tool for future breast cancer therapy? Crit Rev Oncog, 12, 127-50. https://doi.org/10.1615/CritRevOncog.v12.i1-2.70
  13. Tone Y, Toh-E A (2002). Nob1p is required for biogenesis of the 26S proteasome and degraded upon its maturation in Saccharomyces cerevisiae. Genes Dev, 16, 142-57.
  14. Xu G, Bernaudo S, Fu G, et al (2008). Cyclin G2 is degraded through the ubiquitin-proteasome pathway and mediates the antiproliferative effect of activin receptor-like kinase 7. Mol Biol Cell, 19, 4968-79. https://doi.org/10.1091/mbc.E08-03-0259
  15. Zhang Y, Ni J, Zhou J, et al (2005). Cloning, expression and characterization of the human NOB1 gene. Mol Biol Rep, 32, 185-9. https://doi.org/10.1007/s11033-005-3141-7

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