Protein phosphatase inhibitor-1 plays an important role in the regulation of
glycogen metabolism through inhibition of
protein phosphatase-1 activity, and it has been implicated in the regulation of cell growth. Using real-time quantitative RT-PCR, we studied the
mRNA expression of inhibitor-1 in
hepatocellular carcinomas induced in rats by
oral administration of
N-nitrosomorpholine, and in a non-tumorigenic liver cell line (C1I), that stores
glycogen in excess during early passages. In late passages,
glycogen is gradually lost concomitant with cell transformation. Our in vitro model included a tumorigenic subline of C1I cells that was obtained by chemically-induced neoplastic transformation using
N-methyl-N'-nitro-N-nitrosoguanidine (C1Ict), and does not store
glycogen, as well as
Morris hepatoma 3924A (MH3924A) cells. We found that in
hepatocellular carcinomas, in the late
glycogen-poor passages (C1I(late)), and in the tumorigenic subline (C1Ict) of C1I cells, and in MH3924A cells the
mRNA expression of inhibitor-1 is significantly increased. This increase in expression varied from 15 to 290-fold of that observed in normal liver. In contrast, in the early
glycogen-storing passage of C1I cells (C1I(early)) the level of inhibitor-1
mRNA was found to be slightly less than that of normal liver. Inhibitor-1
mRNA levels correlated with the degree of differentiation of HCCs. These results indicate that the expression of inhibitor-1
mRNA is tightly linked to
tumor progression and to the process of liver cell transformation in vitro and is inversely correlated with the
glycogen content of the cell.