ANALYSIS OF CELL GROWTH AND TRANSFORMATION BY ORNITHINE DECARBOXYLASE

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In order to identify gene differentially expressed by putrescine, a polyamine, which play important roles in the regulation of cell proliferation and the development of cancer, we performed mRNA differential display analysis using total RNA extracted from HepG2 cells (human hepatoblastoma cell line) treated with a specific inhibitor of polyamine biosynthesis, α-difluoromethyl ornithine (DFMO). A total of 25 genes were up-regulated and 32 genes down-regulated by putrescine. Of the genes differentially expressed by putrescine, we chose three that were related to the respiratory chain and oxidative phosphorylase and analyzed them by Northern blot analysis. Cytichrome oxidase submit 1, low molecular mass ubiquinone- binding protein, and NADH dehydrogenase subunit 2 were found to down- regulated by putrescine. We examined intracellular ATP level in Hep G2 cells, and found that ATP level in DFMO- treated cells was increased by exogenous putrescine.