Qin Lu,Yang Yixuan,Yan Ning,Chen Yangmei,Chen Lifen.Application of iTRAQ in screening differentially expressed proteins in children with cortical dysplasia-induced epilepsy[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(14):1400-1405.
Qin Lu,Yang Yixuan,Yan Ning,Chen Yangmei,Chen Lifen.Application of iTRAQ in screening differentially expressed proteins in children with cortical dysplasia-induced epilepsy[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(14):1400-1405.DOI:
Application of iTRAQ in screening differentially expressed proteins in children with cortical dysplasia-induced epilepsy
Objective To screen the differentially expressed proteins in children with disorders of cortical development (DCD) accompanied by epilepsy. Methods The brain tissues from children with DCD-induced epilepsy and normal children (control group) operated for head trauma were selected as specimens. Isobaric tags for relative and absolute quantitation (iTRAQ) technique and 2-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) were used for identifying the differentially expressed proteins. The differentially expressed proteins were analyzed by bioinformatics for searching candidate proteins. Finally
the candidate proteins were verified by real-time PCR and Western blotting. Results A total of 153 proteins showed significantly different expression
and included 64 up-regulated proteins and 89 down-regulated proteins. Gene ontology (GO) functional analysis showed that the differentially expressed proteins were mainly involved in catalytic activity. The protein interaction network showed that transferrin (TF)
apolipoprotein A1 (APOA1)
alpha-2-macroglobulin (A2M)
ATPase (ATP6V1A)
and vimentin (VIM) were located at the crossed nodes. The expression levels of TF
APOA1
A2M
ATP6V1A and VIM were verified by real-time PCR and Western blotting
and the results were consistent with those of the quantitative mass spectrometry (P0.05). Conclusion Differentially expressed proteins in the children with DCD-induced epilepsy can be effectively screened by iTRAQ technique combined with 2D-LC-MS/MS.