Difference in Protein
Expression in Vero Cells after Antisense-blocking Genes Involved in the
Suppression of Non-targeted Mutagenesis
JIN Jing-Hua, YU Ying-Nian*, QIAN Yu-Li
( Department of Pathophysiology, Medical School of Zhejiang University,
Hangzhou 310031, China )
Abstract 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.
Key words two-dimensional gel
electrophoresis； proteome； differential expression； mutation； non-targeted
*Corresponding author： Tel,86-571-87217149； Fax, 86-571-87217149； e-mail, [email protected]