title: Secreted Opisthorchis viverrini glutathione S-transferase regulates cell proliferation through AKT and ERK pathways in cholangiocarcinoma.
abstract: Opisthorchis viverrini can develop mitogenic substances into the excretory/secretory product (ESP) that may play an important role in promoting the genesis of cholangiocarcinoma (CCA). In the present study, glutathione S-transferase (GST) is identified as being secreted into Ov-ESP and acting as one of the parasitic mitogens. Its proliferative effect and possible mechanism were explored and its association with the tumor development is proposed. Ov-ESP was concentrated and purified by gel filtration chromatography. SDS-PAGE, 2-DE, and LC-MS/MS identified GST predominantly expressed in the proliferative ESP fraction. The recombinant OvGST (rOvGST) was produced by wheat germ cell-free expression and confirmed by an MTS assay to have a proliferative function on NIH-3T3 murine fibroblasts and MMNK1 non-tumorigenic human bile duct epithelial cells in a dose dependent manner with different optimal doses. The cell surface binding of rOvGST was confirmed in vitro and the activation of both pAKT and pERK was revealed as the mechanism of OvGST-mediated cell proliferation. With support from the observation of secreted OvGST on the biliary cells surrounding the parasites, it is suggested that OvGST can promote cell proliferation that consequently may accelerate the genesis of CCA.
authors.affiliation: Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Daoyot
authors.initials: D
authors.lastname: Daorueang
authors.forename: Peti
authors.initials: P
authors.lastname: Thuwajit
authors.forename: Sittiruk
authors.initials: S
authors.lastname: Roitrakul
authors.forename: Thewarach
authors.initials: T
authors.lastname: Laha
authors.forename: Sasithorn
authors.initials: S
authors.lastname: Kaewkes
authors.forename: Yaeta
authors.initials: Y
authors.lastname: Endo
authors.forename: Chanitra
authors.initials: C
authors.lastname: Thuwajit
mesh_terms: D000818:Animals; D001650:Bile Duct Neoplasms / Q000378:metabolism* / Q000469:parasitology / Q000503:physiopathology; D001653:Bile Ducts, Intrahepatic / Q000378:metabolism / Q000473:pathology; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D018281:Cholangiocarcinoma / Q000378:metabolism* / Q000469:parasitology / Q000473:pathology; D002853:Chromatography, Liquid; D006224:Cricetinae; D003530:Cyprinidae / Q000469:parasitology; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism*; D015801:Helminth Proteins / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D008297:Male; D058488:Metacercariae / Q000737:chemistry / Q000254:growth & development / Q000276:immunology; D051379:Mice; D041681:NIH 3T3 Cells; D009889:Opisthorchiasis / Q000150:complications / Q000378:metabolism* / Q000469:parasitology / Q000473:pathology; D009891:Opisthorchis / Q000737:chemistry* / Q000254:growth & development / Q000276:immunology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D013785:Thailand
chemical_list: D015801:Helminth Proteins; D005982:Glutathione Transferase; D051057:Proto-Oncogene Proteins c-akt