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Abstract
Expression of Cyp1a1 and Cyp1a2 genes was investigated in adult C57BL/6NCrj mouse hepatocytes in primary culture for up to 5 days. When the cells were cultivated as monolayers on collagen‐coated dishes, CYP1A1 mRNA species were prominently induced after treatment with 3‐methylcholanthrene (MCA) throughout the observation period. Substantial induction of CYP1A2 mRNA by MCA was also observed at day 1 of cultivation, followed by a decrease to very low levels thereafter. In contrast, when cultivated on non‐coated dishes, the hepatocytes formed multicellular aggregates (spheroids) and prominent induction of both mRNA species was found for up to 5 days. Constitutive expression of CYP1A2 mRNA in spheroid culture was maintained throughout the observation period, whereas that in monolayer culture decreased rapidly. The time‐course of the induced CYP1A2 mRNA amounts after the treatment with MCA or 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) followed the same pattern as that of CYP1A1 mRNA. Expressed amounts of CYP1A1 or CYP1A2 mRNA in spheroid culture were higher than or similar to the levels in the case of in vivo production, respectively. Induction of both mRNA species was also observed in hepatocytes from nonresponsive DBA/2NCrj mouse in spheroid culture, but the expressed amount after MCA treatment was far smaller than for C57BL/6NCrj cells, despite equivalent expression in the two strains after TCDD. Activities of aryl hydrocarbon hydroxylase (AHH) and acetanilide 4‐hydroxylase (AAH) were elevated with either type of cultivation after treatment with MCA or TCDD, Ratios of AAH to AHH were not changed between the two cultures after 24 h treatment. However, the ratios in spheroid culture after 48 h treatment increased, whereas they did not change in monolayer culture. The present observations indicate that the spheroid culture is more suitable than the monolayer system for studying the mechanism of Cyp1a2 gene expression in adult mouse hepatocytes.
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