title: Serum protein N-glycan alterations of diethylnitrosamine-induced hepatocellular carcinoma mice and their evolution after inhibition of the placental growth factor.
abstract: Placental growth factor (PlGF) inhibition produced promising results in reducing tumor burden in a diethylnitrosamine (DEN)-induced mouse model for hepatocellular carcinoma (HCC). The aim of this study was to non-invasively assess the improved histology by performing a serum glycomic analysis. To elucidate the molecular mechanism underlying the observed glycomic effects, we investigated the transcription and expression of E26 transformation-specific sequence 1 (Ets-1), a transcription factor essential for the glycomic and angiogenic changes in malignant transformation, including its different phosphorylated forms that result from activation of the MAP kinase and a Ca(2+)-dependent pathway. In addition, three Ets-1-dependent glycosyltransferase genes, Mgat4a, Mgat4b, and Mgat5, were also evaluated. HCC was induced in mice by weekly injections with DEN for 16, 20, 25, and 30 w. In the treatment study, mice were injected with DEN for 25 w and subsequently treated with PlGF antibodies (5D11D4) for 5 w. Finally, PlGF-/- mice were injected with DEN for 20, 25, and 30 w. Serum N-glycans were analyzed with DNA sequencer-assisted fluorophore-assisted capillary electrophoresis and compared with histology. Maximum altered N-glycan phenotype was reached after 20 w of DEN-injections, i.e., when the first neoplastic lesions started to appear. 5D11D4-treatment improved the glycomic phenotype in that 7 of the 11 altered glycans tended to normalize. The PlGF-/- mice also showed a normalization trend, although not to the same extent of the treatment group. Number of Ets1, Mgat4a, Mgat4b, and Mgat5 transcripts increased considerably in DEN-injected mice, however, a non-significant decrease was observed after 5D11D4-treatment. On the protein level, 5D11D4-treatment had a prominent effect on the MAP kinase pathway with a significant p38 activation, yet independent of Ets-1 function.
authors.affiliation: Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium. bram.blomme@ugent.be
authors.forename: Bram
authors.initials: B
authors.lastname: Blomme
authors.forename: Femke
authors.initials: F
authors.lastname: Heindryckx
authors.forename: Jean Marie
authors.initials: JM
authors.lastname: Stassen
authors.forename: Anja
authors.initials: A
authors.lastname: Geerts
authors.forename: Isabelle
authors.initials: I
authors.lastname: Colle
authors.forename: Hans
authors.initials: H
authors.lastname: Van Vlierberghe
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000008:administration & dosage; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001798:Blood Proteins / Q000378:metabolism*; D020013:Calcium Signaling; D006528:Carcinoma, Hepatocellular / Q000097:blood* / Q000139:chemically induced / Q000378:metabolism; D002471:Cell Transformation, Neoplastic / Q000378:metabolism; D004052:Diethylnitrosamine; D006031:Glycosylation; D008114:Liver Neoplasms, Experimental / Q000097:blood* / Q000139:chemically induced / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D018345:Mice, Knockout; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D000072483:Placenta Growth Factor; D011134:Polysaccharides / Q000097:blood; D011257:Pregnancy Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D050784:Proto-Oncogene Protein c-ets-1 / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000970:Antineoplastic Agents; D001798:Blood Proteins; C494042:Ets1 protein, mouse; C000608037:Pgf protein, mouse; D011134:Polysaccharides; D011257:Pregnancy Proteins; D050784:Proto-Oncogene Protein c-ets-1; D000072483:Placenta Growth Factor; D004052:Diethylnitrosamine; D017351:N-Acetylglucosaminyltransferases