DOI QR코드

DOI QR Code

Silencing of the COPS3 Gene by siRNA Reduces Proliferation of Lung Cancer Cells Most Likely via induction of Cell Cycle Arrest and Apoptosis

  • Wang, Xue-Mei (Cancer Center, the First Hospital of Jilin University) ;
  • Cui, Jiu-Wei (Cancer Center, the First Hospital of Jilin University) ;
  • Li, Wei (Cancer Center, the First Hospital of Jilin University) ;
  • Cai, Lu (Cancer Center, the First Hospital of Jilin University) ;
  • Song, Wei (Cancer Center, the First Hospital of Jilin University) ;
  • Wang, Guan-Jun (Cancer Center, the First Hospital of Jilin University)
  • 발행 : 2012.03.31

초록

The COPS3 gene has stimulating effect on cell proliferation and progression of osteosarcomas and related cells. However, the features of COPS3 and its potential application as a therapeutic target in other cancers has not yet been studied. In this study, therefore, the effect of COPS3 silencing via COPS3 siRNA on lung cancer cell proliferation was examined. Expression levels of COPS3 gene in COPS3 siRNA infected cells and control siRNA infected cells were compared with real time PCR and Western blot analysis. Cell proliferation levels were comprehensively analyzed by MTT, BrdU incorporationy, and colony formation assays. For mechanistic assessment the effects of COPS3 silencing on cell cycle and apoptosis were analyzed using flow cytometry. Results showed that successful silencing of the COPS3 gene at both translational and transcriptional levels significantly reduced the proliferation and colony formation by lung cancer cells (p<0.01). Flow cytometry showed cell cycle arrest in the G0/G1 phase after COPS3 silencing, and more importantly, apoptosis was induced as a result of COPS3 knockdown, which negatively affected cell survival. Therefore, these results provide another piece of important evidence that the COPS3 gene expressed in lung cancer cells may play a critical role in stimulating proliferation. Down-regulation of COPS3 could significantly inhibit lung cancer cell growth, which was most likely mediated via induction of cell cycle arrest in G0/G1 phase and apoptosis.

키워드

참고문헌

  1. Bech-Otschir D, Kraft R, Huang X, et al (2001). COP9 signalosome-specific phosphorylation targets p53 to degradation by the ubiquitin system. EMBO J, 20, 1630-9. https://doi.org/10.1093/emboj/20.7.1630
  2. Broker LE, Giaccone G (2002). The role of new agents in the treatment of non-small cell lung cancer. Eur J Cancer, 38, 2347-61. https://doi.org/10.1016/S0959-8049(02)00457-4
  3. Chen L, Liang Z, Tian Q, et al (2011). Overexpression of LCMR1 is significantly associated with clinical stage in human NSCLC. J Exp Clin Cancer Res, 30, 18. https://doi.org/10.1186/1756-9966-30-18
  4. Devi G R (2006). siRNA-based approaches in cancer therapy. Cancer Gene Ther, 13, 819-29. https://doi.org/10.1038/sj.cgt.7700931
  5. Fuchs B, Pritchard DJ (2002). Etiology of osteosarcoma. Clin Orthop, 397, 40-52. https://doi.org/10.1097/00003086-200204000-00007
  6. Gokgoz N, Wunder JS, Mousses S, et al (2001). Comparison of p53 mutations in patients with localized osteosarcoma and metastatic osteosarcoma. Cancer, 92, 2181-9. https://doi.org/10.1002/1097-0142(20011015)92:8<2181::AID-CNCR1561>3.0.CO;2-3
  7. Henriksen J, Aagesen TH, Maelandsmo GM, et al (2003). Amplification and overexpression of COPS3 in osteosarcomas potentially target TP53 for proteasomemediated degradation. Oncogene, 22, 5358-61. https://doi.org/10.1038/sj.onc.1206671
  8. Luo J, Emanuele MJ, Li D, et al (2009). A genome-wide RNAi screen identifies multiple synthetic lethal interactions with the Ras oncogene. Cell, 137, 835-48. https://doi.org/10.1016/j.cell.2009.05.006
  9. Paddison PJ, Hannon GJ (2003). siRNAs and shRNAs: skeleton keys to the human genome. Curr Opin Mol Ther, 5, 217-24.
  10. Sharma SV, Bell DW, Settleman J, Haber DA (2007). Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer, 7, 169-81. https://doi.org/10.1038/nrc2088
  11. Singh A, Boldin-Adamsky S, Thimmulappa RK, et al (2008). RNAi-mediated silencing of nuclear factor erythroid-2- related factor 2 gene expression in non-small cell lung cancer inhibits tumor growth and increases efficacy of chemotherapy. Cancer Res, 68, 7975-84. https://doi.org/10.1158/0008-5472.CAN-08-1401
  12. Skrzypski M, Dziadziuszko R, Jassem J (2011). MicroRNA in lung cancer diagnostics and treatment. Mutat Res, 17, 25-31.
  13. van Dartel M, Redeker S, Bras J, Kool M, Hulsebos TJ (2004). Overexpression through amplification of genes in chromosome region 17p11.2 approximately p12 in highgrade osteosarcoma. Cancer Genet Cytogenet, 152, 8-14. https://doi.org/10.1016/j.cancergencyto.2003.09.024
  14. Wei N, Deng XW (2003). The COP9 signalosome. Annu Rev Cell Dev Biol, 19, 261-86. https://doi.org/10.1146/annurev.cellbio.19.111301.112449
  15. Yan J, Walz K, Nakamura H, et al (2003). COP9 signalosome subunit 3 is essential for maintenance of cell proliferation in the mouse embryonic epiblast. Mol Cell Biol, 23, 6798-808. https://doi.org/10.1128/MCB.23.19.6798-6808.2003
  16. Yan T, Wunder JS, Gokgoz N, et al (2007). COPS3 amplification and clinical outcome in osteosarcoma. Cancer, 109, 1870-6. https://doi.org/10.1002/cncr.22595
  17. Yan T, Tang G, Ren T, et al (2011). RNAi-mediated COPS3 gene silencing inhibits metastasis of osteogenic sarcoma cells. Cancer Gene Ther, 18, 450-6. https://doi.org/10.1038/cgt.2011.16

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