Chinese Clinical Oncology ›› 2018, Vol. 23 ›› Issue (6): 508-513.
Previous Articles Next Articles
Online:
Published:
Abstract: ObjectiveTo investigate the effect of RNA interference on UHRF1 gene expression and proliferation and apoptosis of glioma U251 cells. MethodsThe plasmid LVUHRF1shRNA and negative control plasmid LVscrambleshRNA were transfected into U251 cells (LVUHRF1shRNA group and LVscrambleshRNA group) using lipid Attractene reagent, respectively. Cells only transfected liposomes were used as the control group. The UHRF1 mRNA level was detected by realtime 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 VFITC/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, caspase3 and Bcl2) at 72 h after transfection. ResultsThe results of QPCR test showed that the level of UHRF1 in the control group, the LVscrambleshRNA group and the LVUHRF1shRNA group were 1098±0136, 1127±0193 and 0309±0073, respectively. The UHRF1 levels in the LVUHRF1shRNA group was lower than those of the control group and the LVscrambleshRNA group (P<005). Compared with the control group and the LVscrambleshRNA group, the cell proliferation rate and the BrdU positive rate were decreased but the apoptosis rate increased in the LVUHRF1shRNA group (P<005). The apoptotic rate was (2571±187)% in LVUHRF1shRNA group, higher than (773±066)% of the control group and (786±059)% of the LVscrambleshRNA group, and the difference was statistically significant (P<005). The results of Western blotting showed that compared with the control group and the LVscrambleshRNA group, the level of Bax and caspase3 increased but the Bcl2 level decreased in the LVscrambleshRNA group, and the difference was statistically significant (P<005). There was no significant difference in the above indices between the control group and the LVscrambleshRNA group (P>005). ConclusionThe downregulation 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
ZHANG Yong, DING Lianshu. . Effects of RNA interference on UHRF1 gene expression and proliferation and apoptosis of glioma U251 cells#br#
0 / / Recommend
Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks
URL: http://manu65.magtech.com.cn/Jwk3_lczlxzz/EN/
http://manu65.magtech.com.cn/Jwk3_lczlxzz/EN/Y2018/V23/I6/508
Cited