Mechanism of action of ethanol in the down-regulation of Gal(beta)1, 4GlcNAc alpha2,6-sialyltransferase messenger RNA in human liver cell lines

• Published in: Metabolism, clinical and experimental Vol. 54; no. 6; pp. 729 - 734
• Main Authors: Garige, Mamatha, Gong, Maokai, Rao, Manjunath N, Zhang, Yili, Lakshman, M Raj
• Format: Journal Article
• Language: English
• Published: United States 01-06-2005
• Subjects: Alcohol Dehydrogenase - genetics; Cell Line; Cytochrome P-450 CYP2E1 - genetics; Down-Regulation; Ethanol - pharmacology; Gene Expression Regulation, Enzymologic - drug effects; Humans; Liver - drug effects; Liver - enzymology; RNA, Messenger - analysis; Sialyltransferases - genetics
• ISSN: 0026-0495

Gal beta l, 4GlcNAc alpha 2,6-sialyltransferase (2,6-ST) mediates the addition of alpha 2,6-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 micro mol/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.