The Study On The Correlation Between Notch-1Gene And Autophagy In Human Glioma Cell

Objective:To investigate autophagy phenomenon in human glioma cell when the expressionof Notch1gene were down-regulated or up-regulated and research the possiblemechanism of Notch-1gene involved in autophagy in human glioma cell.Methods:The lentiviral vectors, which expressed Notch1-shRNA or Notch-1intracellulardomain (NICD), were transfected into human glioma U251cell respectively. RT-PCRand Western blotting were applied to monitor the validity of down-regulation ofNotch1expression and over-expression of NICD. MDC staining was performed toobserve the cell autophagy. Western blotting and immunofluorescence staining wereapplied to monitor the expression of the autophagy related protein LC3(microtubule-associated protein1light chain3) and Beclin-1.Results:The lentiviral vectors, which expressed Notch1-shRNA or NICD, were efficientin down-regulation of Notch-1expression and over-expression of NICD respectively.MDC staining revealed that punctuate autophagic vacuoles (AVs) were appeared inglioma U251cells which Notch-1gene was knockdown. Western blotting andimmunofluorescence staining showed that the expression of the autophagy relatedprotein LC3and Beclin-1increased in glioma U251cells when the expression ofNotch1gene were down-regulated.Conclusions:Autophagy phenomenon can be observed in human glioma cells when the expression of Notch-1gene was down-regulated. The autophagy associated withNotch-1knockdown in glioma cells may be induced by up-regulating the expressionsof LC-3and Beclin-1.