Chinese Journal of Contemporary Neurology and Neurosurgery (Jul 2013)
Expression profiles of genes in wild-type DJ-1 and L10P mutant DJ-1 in monoclonal cell strains
Abstract
Background DJ⁃1 gene is a causative gene which contributes to the onset of autosomal recessive early-onset parkinsonism (AREP). Many research suggest that DJ-1 protein may change expression of certain genes through regulate its transcriptional activity, which play a role in the pathogenesis of Parkinson's disease (PD). In our previous study, we found a new mutation of DJ-1 which we named as L10P. DJ-1 gene encodes the first frame 29 bp from the thymine (T)→cytosine (C), so that the leucine on the 10th locus of DJ-1 protein was replaced by proline (L10P). To elucidate the effect of the L10P mutation, we identify genes for which expressions are abnormally regulated by L10P mutant DJ-1 protein using DNA microarray analysis. Methods Human embryonic kidney cell 293 (HEK293) monoclonal cell strains which can stably express pCMV-Tag2A-Flag, pCMV-Tag2A-Flag-DJ-1 and pCMV-Tag2A-Flag-DJ-1-L10P were selected by screening, and identified on the basis of DNA, RNA and protein levels to confirm whether the acquired HEK293 monoclonal cell strains can stably express empty vector, wild-type DJ-1 protein and L10P mutant DJ-1 protein. Gene chip technique was used to perform differential gene screening for different groups of HEK293 monoclonal cell strains. Results Compared with the expression in empty vector group, the expression of 14 genes was up-regulated and 28 genes was down-regulated in wild-type group; and the expression of 14 genes was up⁃regulated and 9 down-regulated in expressing L10P mutant group respectively. Comparison of the expression in wild⁃type group, expression of 59 genes was up⁃regulated and 27 genes down-regulated in L10P mutant group. These differential genes all took part in the biological processes including signal transduction, transcriptional regulation, cell cycle, apoptosis, oxidative stress and so on. Conclusion L10P mutant DJ-1 protein may directly or indirectly influence the singal transduction and play a role in the mechanism of PD.
