title: Suppression of Ov-grn-1 encoding granulin of Opisthorchis viverrini inhibits proliferation of biliary epithelial cells.
abstract: Multistep processes likely underlie cholangiocarcinogenesis induced by chronic infection with the fish-borne liver fluke, Opisthorchis viverrini. One process appears to be cellular proliferation of the host bile duct epithelia driven by excretory-secretory (ES) products of this pathogen. Specifically, the secreted growth factor Ov-GRN-1, a liver fluke granulin, is a prominent component of ES and a known driver of hyper-proliferation of cultured human and mouse cells in vitro. We show potent hyper-proliferation of human cholangiocytes induced by low nanomolar levels of recombinant Ov-GRN-1 and similar growth produced by low microgram concentrations of ES products and soluble lysates of the adult worm. To further explore the influence of Ov-GRN-1 on the flukes and the host cells, expression of Ov-grn-1 was repressed using RNA interference. Expression of Ov-grn-1 was suppressed by 95% by day 3 and by ~100% by day 7. Co-culture of Ov-grn-1 suppressed flukes with human cholangiocyte (H-69) or human cholangiocarcinoma (KKU-M214) cell lines retarded cell hyper-proliferation by 25% and 92%, respectively. Intriguingly, flukes in which expression of Ov-grn-1 was repressed were less viable in culture, suggesting that Ov-GRN-1 is an essential growth factor for survival of the adult stage of O. viverrini, at least in vitro. To summarize, specific knock down of Ov-grn-1 reduced in vitro survival and capacity of ES products to drive host cell proliferation. These findings may help to contribute to a deeper understanding of liver fluke induced cholangiocarcinogenesis.
authors.affiliation: Graduate School, Khon Kaen University, 40002 Khon Kaen, Thailand; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Atiroch
authors.initials: A
authors.lastname: Papatpremsiri
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, Queensland Tropical Health Alliance Laboratory, James Cook University, Cairns, Queensland 4878, Australia.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Smout
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, Queensland Tropical Health Alliance Laboratory, James Cook University, Cairns, Queensland 4878, Australia.
authors.forename: Alex
authors.initials: A
authors.lastname: Loukas
authors.affiliation: Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Brindley
authors.affiliation: Tropical Disease Research Laboratory, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Banchob
authors.initials: B
authors.lastname: Sripa
authors.affiliation: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Electronic address: thewa_la@kku.ac.th.
authors.forename: Thewarach
authors.initials: T
authors.lastname: Laha
mesh_terms: D000818:Animals; D001650:Bile Duct Neoplasms / Q000469:parasitology / Q000473:pathology / Q000517:prevention & control; D001652:Bile Ducts / Q000166:cytology*; D001653:Bile Ducts, Intrahepatic / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018281:Cholangiocarcinoma / Q000469:parasitology / Q000473:pathology / Q000517:prevention & control; D006224:Cricetinae; D004847:Epithelial Cells / Q000166:cytology; D005786:Gene Expression Regulation; D020868:Gene Silencing; D000077155:Granulins; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008647:Mesocricetus; D009889:Opisthorchiasis / Q000150:complications; D009891:Opisthorchis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000096:biosynthesis / Q000378:metabolism; D018510:RNA, Helminth / Q000302:isolation & purification
chemical_list: D000077155:Granulins; D036341:Intercellular Signaling Peptides and Proteins; D012330:RNA, Double-Stranded; D018510:RNA, Helminth
keywords: Cellular proliferation; Cholangiocarcinoma; Granulin; Liver fluke; O. viverrini; RNA interference