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 non-tumor and tumor samples for proteins with altered abundance in gastric adenocarcinoma (GA). 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- subunit (HADHA) was decreased, and the MTP- subunit (HADHB) was increased. On the other hand, heterogenous nuclear ribonucleoprotein M (HNRNPM) and galectin-4 (LGALS4) were increased in the gastric tumor samples both in 2DE and immunoblotting.
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