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J. Biol. Chem., Vol. 262, Issue 3, 1092-1097, Jan, 1987
JR Gum, WK Kam, JC Byrd, JW Hicks, MH Sleisenger and YS Kim
We have examined the effects of the "differentiating agent," sodium
butyrate, on the induction of alkaline phosphatase in human colonic tumor
cell line LS174T. Culture of these cells in the presence of 2 mM butyrate
caused this activity to increase from less than 0.0001 unit/mg of protein
to greater than 0.7 unit/mg of protein over an 8-day period. This induction
proceeded in a nonlinear fashion with a lag time of 2-3 days occurring
before enzymatic activity began to rise. These increases in activity were
accompanied by elevations in the content of a placental-like isozyme of
alkaline phosphatase as demonstrated by "Western" immunoblots. Dome
formation, indicative of differentiation in cultured cells, also required 3
days treatment with butyrate before becoming evident. The rate of
biosynthesis of the enzyme, examined using metabolic labeling with
L-[35S]methionine and immunoprecipitation, was found to increase
continuously between days 2 and 6 of butyrate treatment. "Northern" blot
analysis indicated that treatment of these cells with butyrate caused
greater than 20-fold induction of a 2700-base mRNA that hybridized to a
cDNA probe for placental alkaline phosphatase. The mRNA for alkaline
phosphatase produced by these cells upon butyrate treatment was
approximately 300- 400 bases smaller than the mRNA for alkaline phosphatase
found in placenta. Human small intestine also contained two mRNAs that
hybridized relatively weakly with the placental alkaline phosphatase probe.
These results indicate that a placental alkaline phosphatase- like protein
and mRNA are induced by butyrate in LS174T cells with a time course
consistent with cellular differentiation preceding induction.
Effects of sodium butyrate on human colonic adenocarcinoma cells. Induction of placental-like alkaline phosphatase
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