2DE in combination with MS has facilitated the discovery of several
proteins with altered abundance in
gastric cancer. While acidic and wide pH ranges have been widely investigated, analysis in the alkaline pH range has not been specifically performed in
gastric cancer to date. In the present study, we initially optimized the 2DE in alkaline pH range (pH 7-11) for gastric tissue samples. Using a modified lysis
buffer, we analyzed pooled nontumor and
tumor samples for
proteins with altered abundance in gastric
adenocarcinoma. We successfully identified 38
silver-stained spots as 24 different
proteins. Four of these were chosen for investigation with immunoblotting on individual paired samples to determine whether the changes seen in 2DE represent the overall abundance of the
protein or possibly only a single form. While
mitochondrial trifunctional protein (MTP) subunits were decreased in 2DE
gels, immunoblotting identified their overall abundance as being differently dysregulated: in the gastric
tumor samples, the MTP-α subunit was decreased, and the MTP-β subunit was increased. On the other hand, heterogenous nuclear
ribonucleoprotein M and
galectin-4 were increased in the gastric
tumor samples in both 2DE and immunoblotting.