Inactivation of phosphatidylethanolamine N-methyltransferase-2 in aflatoxin-induced liver cancer and partial reversion of the neoplastic phenotype by PEMT transfection of hepatoma cells.
Phosphatidylethanolamine N-methyltransferase(PEMT) is an enzyme in liver that catalyzes the stepwise methylation of phosphatidylethanolamine to phosphatidylcholine, in addition to the main pathway that synthesizes phosphatidylcholine directly from choline. We have reported that PEMT is permanently inactivated in liver cancer induced by the Solt and Farber model. Here we studied, (i) whether similar changes also occur in the progression of hepatocarcinoma triggered by aflatoxin B(1) (AFB(1)) in rats; (ii) whether the hepatoma phenotype could be reversed by over-expression of PEMT2. We found that PEMT2 protein decreased in pre-neoplastic nodules and virtually disappeared in hepatocellular carcinoma induced by AFB(1) due to decreased levels of mRNA without any deletion or mutation of the DNA sequence. PEMT activity, which reflects the function of both PEMT1 and PEMT2, was lower in nodules and negligible in the tumor, consistent with its regulation at the level of gene transcription. McArdle hepatoma cells transfected with PEMT2 failed to form anchorage-independent colonies in soft agar, while the vector-transfected control line grew efficiently. Moreover, PEMT2-transfected cells were also poorly tumorigenic in vivo in athymic mice, as shown by the lower tumor incidence, the longer cancer-free-time and the lower tumor volume and weight. Together, these data indicate that the loss of PEMT function may contribute to malignant transformation of hepatocytes.
Tessitore
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Department of Medical Sciences, "Amedeo Avogadro" University of East Piedmont, Turin, Italy. tessitor@pasteur.sluigi.unito.it
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D000348:Aflatoxins / Q000633:toxicity; D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced / Q000235:genetics / Q000473:pathology / Q000628:therapy; D008297:Male; D008780:Methyltransferases / Q000235:genetics*; D051379:Mice; D050918:Phosphatidylethanolamine N-Methyltransferase; D051381:Rats; D017208:Rats, Wistar
D000348:Aflatoxins; D008780:Methyltransferases; D050918:Phosphatidylethanolamine N-Methyltransferase