Abstract
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.
Highlights
Lung cancer is one of the major causes of human death both in Asian and Western populations and has become a heavy burden disease in the world wide (Sharma et al, 2007)
The protein expression levels were analyzed by Western blotting, which demonstrated that the COPS3 expression is detected by Western blotting in both A549 and 95D lung cancer cells in control SiRNA group, and significant suppression of COPS3 protein expression was evident in cells treated with specific small interfering RNA (siRNA) compared to control siRNA (Figure 1C)
In the present study efforts have been taken to identify the efficiency of COPS3 gene silencing through siRNA technology as a therapeutic target in treatment of lung cancer cells
Summary
Lung cancer is one of the major causes of human death both in Asian and Western populations and has become a heavy burden disease in the world wide (Sharma et al, 2007). The number of lung cancer patients are increasing annually. An exciting report by Singh et al showed that siRNA-mediated silencing of nuclear factor erythroid-2–related factor 2 gene expression in non-small cell lung cancer inhibited tumor growth and increased efficacy of chemotherapy (Singh et al, 2008). Identification and silencing of genes related to the growth and survival of lung cancer cells would be an interesting area of research. To this end, lentivirus based short hairpin RNA (shRNA) is widely used to target specific gene in cancer cells (Luo et al, 2009)
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