Résumé
Objective To explore the effect of 14-3-3 β gene on biological behavior ofglioma cell line and its mechanism.Methods Conventional cultured SVGp12,U251,U87 and SHG-44 cell lines and U251 cells silenced by 14-3-3[β-small interfering RNA (siRNA) were collected; real time-PCR and Western blotting were used to detect the 14-3-3β gene and protein expressions in these cells.Conventional cultured U251 cells at logarithmic phase were divided into three groups:experimental group (14-3-3β-siRNA transfection),negative control group (siRNA transfection) and blank control group; 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay was used to assess the proliferation of U251 cells,flow cytometry was used to test the cell apoptosis,and cell migration was analyzed by Transwell chamber assay.Results As compared with those in the normal glial cells,14-3-3β gene and protein expression levels in the glioma cells were significantly higher (P＜0.05); as compared with negative control and blank control groups,U251 cells in the experimental group had significantly decreased gene and protein expressions of 14-3-3β,decreased proliferation and migration abilities,significantly increased apoptosis rate and p53 mRNA level (P＜0.05).Conclusion Silence of 14-3-3 β gene decreases U251 cells proliferation and migration through p53 mediated pathway; consequently,a new explanation about how 14-3-3 β regulates glioma cells proliferation and migration can be clarified,and a potential target for glioma treatment can be provided.
Résumé
There are two kinds of treatments on Alzheimer's disease (AD) rat by transplanting neural stem cells (NSCs),i.e.the replacing cell curing and the gene therapy.By replacing method,the AD rats showed signs of recovering to some extent on both histomorphology and behavior after transplanting NSCs into their brains.Transplanting NSCs along with the nerve nutrition factor (NTFs) showed better curative effects than NSCs transplantation alone.However,little is known about the molecular mechanism involving in the development of NSCs in vivo conditions.And the blindness of the treatment hindered the comparison of various affecting factors.The NSCs gene therapy is still in initial studying,with the effects of both cell replacement and gene therapy.This treatment genetically modified NSCs mainly by unitary nutrition fators such as nerve growth factor (NGF),brain-derived neurotrophic factor (BDNF),and glial cell line-derived neurotrophic factor (GDNF).And it was almost known nothing about the exogenous gene expression efficiency,the induce differentiation,the restore of function and security after genetically-modified NSCs transplanted into the AD rat brain.The detecting technology of NSCs transplanting curative effects of the AD rat is unitary at present.And the combined method is the developing trend,such as combining the immunohistochemical method with in vivo-tracking,and combining morphology index with the function index.