Mechanism of action of ethanol in the down-regulation of Galβ1, 4GlcNAc α2,6-sialyltransferase messenger RNA in human liver cell lines

Galβl, 4GlcNAc α2,6-sialyltransferase (2,6-ST) mediates the addition of α2,6-linked sialic acid to glycoproteins in the Golgi compartment. Down-regulation of its gene and consequent impaired activity of 2,6-ST seems to be the major cause for the appearance of asialoconjugates in the blood of long-term alcoholics. Therefore, mechanism(s) involved in the regulation of 2,6-ST gene is important and clinically relevant. Our previous work showed that long-term ethanol feeding in rats caused a marked 59% decrease of 2,6-ST activity as well as 2,6-ST messenger RNA (mRNA) level in liver that were due to the decreased stability of its mRNA. We now mimic these actions of ethanol using (a) human liver HepG2 cells stably transfected with ethanol-inducible human cytochrome P4502E1 (CYP2E1 cells), or (b) with high alcohol dehydrogenase (HAD cells) but not in wild-type HepG2 cells lacking either of the above 2 enzymes as models. Incubation of these cells for 72 hours with 100 mmol/L ethanol caused decreases (up to 76%, P < .05) of 2,6-ST mRNA levels in CYP2E1 and HAD cells but not in the wild type. However, incubation of wild-type cells with acetaldehyde at concentrations of 50 and 100 μmol/L showed a dramatic decrease (up to 69%, P < .02) in the 2,6-ST mRNA levels. Furthermore, exposure of CYP2E1 cells to 4-hydroxy-2-nonenal, an endogenous lipid peroxidation product of reactive oxygen species, strongly decreased 2,6-ST mRNA level by 61% (P < .02). These results demonstrate that 2,6-ST gene is highly sensitive to ethanol action in human liver cells either via its oxidation product, acetaldehyde, or via reactive oxygen species leading to the generation of a more reactive aldehyde such as 4-hydroxy-2-nonenal. Thus, this study assumes major importance and clinical relevance because 2,6-ST gene regulation in a human liver cell model is demonstrated within a few days of ethanol exposure, whereas its in vivo regulation in liver generally takes prolonged period of ethanol exposure.