Difference in Protein Expression in Vero Cells after Antisense-blocking Genes Involved in the Suppression of Non-targeted Mutagenesis.
A cDNA fragment (fragment 9) has been isolated by mRNA differential display and antisense technology in this lab, and its relevant gene (fragment 9 related gene, FNR gene) might be involved in the inhibition of non-targeted mutagenesis induced by N -methyl- N ' -nitro-N-nitrosoguanidine(MNNG) in mammalian cells. In order to elucidate the functional mechanism of the FNR gene, the protein expression was compared between MNNG-exposed Vero cells transfected with antisense RNA expression plasmid (Vero-pM-amp(-)-9(-)) and those with vector DNA (Vero-pM-amp(-)), by using two-dimensional gel electrophoresis followed by 2D image software analysis. Our analysis indicated that 12 proteins were specifically expressed only in Vero-pM-amp(-)-9(-), and 2 proteins in Vero-pM-amp(-). In addition,there were 24 proteins expressed in higher level in Vero-pM-amp(-)-9(-) as compared with Vero-pM-amp(-)( P <0.05), among them the expression of 7 proteins were enhanced by greater than 5 folds. These results suggest that antisense blocking the FNR gene expression triggered a series of alteration of other gene expression and the FNR gene might be a regulatory factor. This study will also facilitate the identification and characterization of these proteins and corresponding genes involved in the non-targeted mutagenesis.
Jin
J H
JH
Department of Pathophysiology, Medical School of Zhejiang University, Hangzhou 310031, China. ynyu@mail.hz.zj.cn
Yu
Y N
YN
Qian
Y L
YL