Chinese Clinical Oncology ›› 2018, Vol. 23 ›› Issue (6): 508-513.

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Effects of RNA interference on UHRF1 gene expression and proliferation and apoptosis of glioma U251 cells#br#
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  1. Department of Neurosurgery, the First Hospital of Huaian Affiliated to Nanjing Medical University Corresponding author: DING Lianshu
  • Online:2018-06-30 Published:2018-08-29

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

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