Increased phospholipase D activity in butyrate-induced differentiation of HT-29 cells

Phospholipids are important constituents of biomembrane components and are supposed to function as enzyme activators or precursors of bioactive substances. Our earlier work has shown an increased esterification of neutral lipids of HT-29 cells during butyrate-induced differentiation (M. Madesh, O. Benard, K.A. Balasubramanian, Butyrate-induced alteration in lipid composition of human colon cell line HT-29, Biochem. Mol. Biol. Int. 38 (1996) 659-664). In this report we show that there is an increase in phospholipase D (PLD) activity during butyrate-induced differentiation of HT-29 cells as indicated by the formation of phosphatidic acid (PA). When the control and butyrate-treated cell homogenates were incubated in vitro with 1 mM Ca2+, the increase in PA formation was higher than in butyrate-treated cells. This PA was formed due to PLD activity that was confirmed by the generation of phosphatidylethanol by in vitro incubation of HT-29 cell homogenates in the presence of ethanol. The formation of PA was associated with a decrease in phosphatidylcholine (PC) and phosphatidylethanolamine (PE). This study has shown an increase in PLD activity associated with the differentiation of HT-29 cells.