Title: The effect of silencing Dicer by RNA interference on malignant phenotype of glioma cells
Abstract: Objective To investigate the effect of knocking down Dicer by RNAi on the biological characteristics of human glioma cells U251 in vitro.Methods The recombinant adenovirus expression vector which contained short hairpin RNA targeting Dicer (rAd-Dicer) was transfected into U251 cells.The silencing effect of RNAi on Dicer expression was identified by RT- PCR,immumofluorecence staining and Western blot.Western blot was also undertaken to analyze the expression of some functional proteins.The biological behavior changes of U251 cells trasfected by rAd- Dicer were evaluated by MTT,FCM and transwell assay.Methods rAd-Dicer dramatically down- regulated the expression of Dier in U251 cells.As compared with parental and rAd- HK transfected cells,the protein expression of p- AKT,MMP2/9,PCNA,Cyclin a,VEGF,CD34 were up- regulated in rAd-Dicer treated cells.Decreased dicer expression in rAd-Dicer transfected cells was accompanied by 14.1% to 15.3% increase in U251 cells in S phase.Meanwhile,the proliferation and invasive activity were significantly enhanced in rAd-Dicer transfected cells by the MTT and transwell assay.Conclusion Knockdown of dicer renders the U251 cells more malignant transformation.This preliminary finding suggests that global lowering expression of microRNAs may have an oncogenic role.
Key words:
Glioma; microRNA; Dicer; RNA interference; Biological phenotype
Publication Year: 2011
Publication Date: 2011-02-28
Language: en
Type: article
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