Triiodothyronine stimulates the expression of sucrase-isomaltase in Caco-2 cells cultured in serum-free medium

Experimental Cell Research
C JumarieC Malo

Abstract

In a previous study we have shown that triiodothyronine (T3) added to a serum-free medium supplemented with insulin, transferrin, and selenous acid (ITS) can stimulate Caco-2 cell differentiation. In this study we have focused on the effects of T3 on sucrase activity. The results obtained demonstrate that T3 (50 nM) does not change Caco-2 cell proliferation but enhances sucrase activity from 50 to 80%. Similar increases were observed whether or not insulin was present in the culture medium, showing that there was no synergistic effect between T3 and insulin on sucrase activity. Moreover, T3 acts specifically during the differentiation period since addition of T3 to the defined TS medium before confluency is reached does not stimulate sucrase activity. Sucrase kinetic parameters were evaluated for the first time in Caco-2 cells under various culture conditions. The presence of a single enzyme was verified, with a Km of about 7 mM and a Vmax around 20 nmol of substrate hydrolyzed min-1 mg-1 of protein. Our results showed that T3 did not change the enzyme's affinity for sucrose but doubled the Vmax. Moreover, immunoblotting using anti-sucrase-isomaltase (SI) antibodies revealed an approximately twofold increase in the relative amo...Continue Reading

Citations

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