关键词: 脑胶质瘤, UHRF1, 增殖, 凋亡

Abstract: ObjectiveTo investigate the effect of RNA interference on UHRF1 gene expression and proliferation and apoptosis of glioma U251 cells. 
MethodsThe plasmid LVUHRF1shRNA and negative control plasmid LVscrambleshRNA were transfected into U251 cells （LVUHRF1shRNA group and LVscrambleshRNA group） using lipid Attractene reagent， respectively. Cells only transfected liposomes were used as the control group. The UHRF1 mRNA level was detected by realtime fluorescent quantitative PCR （QPCR） at 48 h after transfection. MTT method was used to detect the proliferation of 24， 48 and 72 h in each group. BrdU staining was used to detect the positive rate of BrdU after transfection of 12， 24， 48， 72 h， and Annexin VFITC／PI double staining dye flow cytometry was used to detect the apoptotic rate at 48 and 72 h after transfection. Western blotting was used to detect the levels of apoptosis related proteins （Bax， caspase3 and Bcl2） at 72 h after transfection. 
ResultsThe results of QPCR test showed that the level of UHRF1 in the control group， the LVscrambleshRNA group and the LVUHRF1shRNA group were 1098±0136， 1127±0193 and 0309±0073， respectively. The UHRF1 levels in the LVUHRF1shRNA group was lower than those of the control group and the LVscrambleshRNA group （P＜005）. Compared with the control group and the LVscrambleshRNA group， the cell proliferation rate and the BrdU positive rate were decreased but the apoptosis rate increased in the LVUHRF1shRNA group （P＜005）. The apoptotic rate was （2571±187）％ in LVUHRF1shRNA group， higher than （773±066）％ of the control group and （786±059）％ of the LVscrambleshRNA group， and the difference was statistically significant （P＜005）. The results of Western blotting showed that compared with the control group and the LVscrambleshRNA group， the level of Bax and caspase3 increased but the Bcl2 level decreased in the LVscrambleshRNA group， and the difference was statistically significant （P＜005）. There was no significant difference in the above indices between the control group and the LVscrambleshRNA group （P＞005）. 
ConclusionThe downregulation of UHRF1 in U251 cells can inhibit proliferation and induce apoptosis. UHRF1 plays a role similar to oncogene， which can be used as a candidate gene for the treatment of glioma.

Key words: Glioma, UHRF1, Proliferation, Apoptosis