Journal of Jilin University Medicine Edition ›› 2015, Vol. 41 ›› Issue (04): 701-705.doi: 10.13481/j.1671-587x.20150406

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Inhibitory effect of RNA silencing FZD4 expression on self-renewal ability of glioma stem cells

GAO Xingchun, GUO Na, YAN Aili, FENG Hao, SHI Hongen, GOU Xingchun   

  1. Institute of Basic Medical Science, Xi'an Medical University, Xi'an 710021, China
  • Received:2014-01-12 Published:2015-08-01

Abstract:

Objective To investigate the effect of FZD4 on the self-renewal ability of glioma stem cells,and to clarify its mechanism. Methods Two shRNA sequences were designed according to FZD4 mRNA sequence,named FZD4-shRNA-1# and FZD4-shRNA-2#.Then the lentiviral vector particles were transfected into U251cells(FZD4-shRNA-1# and FZD4-shRNA-2#),The U251 cells infected with the empty vector were used as control group.The expression of green fluorescence protein (GFP) in the transfected cells were observed under inverted fluorescence microscope.The expression levels of FZD4 mRNA and protein in the U251 cells in various gorups were detected by reverse transcription (RT)-PCR and Western blotting method.The U251 cells were cultured in SFM and the glioma stem cells were selected;the proliferation and self-renewal of the tumore stem cells were observed with limiting dilution assay. Results Compared with control group,the expression levels of FZD4 mRNA and protein in FZD4 shRNA groups were significantly reduced(P<0.01).The diameters of neurospheres in FZD4 shRNA groups were significantly smaller than that in control group(P<0.01).The number of cells required to generate at least one tumor sphere/well was significantly increased in FZD4 shRNA groups than that in control group(P<0.05 or P<0.01) in limiting dilution assay. Conclusion Down-regulation of FZD4 expression that in inhibit the self-renewal ability U251 glioma stem cells.

Key words: glioma, frizzled, short hairpin RNA, tumor stem cell, self-renewal

CLC Number: 

  • R739.41