title: Hepatitis B virus PreS2-mutant large surface antigen activates store-operated calcium entry and promotes chromosome instability.
abstract: Hepatitis B virus (HBV) is a driver of hepatocellular carcinoma, and two viral products, X and large surface antigen (LHBS), are viral oncoproteins. During chronic viral infection, immune-escape mutants on the preS2 region of LHBS (preS2-LHBS) are gain-of-function mutations that are linked to preneoplastic ground glass hepatocytes (GGHs) and early disease onset of hepatocellular carcinoma. Here, we show that preS2-LHBS provoked calcium release from the endoplasmic reticulum (ER) and triggered stored-operated calcium entry (SOCE). The activation of SOCE increased ER and plasma membrane (PM) connections, which was linked by ER- resident stromal interaction molecule-1 (STIM1) protein and PM-resident calcium release- activated calcium modulator 1 (Orai1). Persistent activation of SOCE induced centrosome overduplication, aberrant multipolar division, chromosome aneuploidy, anchorage-independent growth, and xenograft tumorigenesis in hepatocytes expressing preS2- LHBS. Chemical inhibitions of SOCE machinery and silencing of STIM1 significantly reduced centrosome numbers, multipolar division, and xenograft tumorigenesis induced by preS2-LHBS. These results provide the first mechanistic link between calcium homeostasis and chromosome instability in hepatocytes carrying preS2-LHBS. Therefore, persistent activation of SOCE represents a novel pathological mechanism in HBV-mediated hepatocarcinogenesis.
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Tim Ting-Chung
authors.initials: TT
authors.lastname: Yen
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Anderson
authors.initials: A
authors.lastname: Yang
authors.affiliation: Department of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan.|Center of Infectious Diseases and Signal Transduction, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Wen-Tai
authors.initials: WT
authors.lastname: Chiu
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Tian-Neng
authors.initials: TN
authors.lastname: Li
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Lyu-Han
authors.initials: LH
authors.lastname: Wang
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Yi-Hsuan
authors.initials: YH
authors.lastname: Wu
authors.affiliation: Institute of Pharmaceutics, Development Center for Biotechnology, Taipei 22180, Taiwan.
authors.forename: Hui-Chen
authors.initials: HC
authors.lastname: Wang
authors.affiliation: Institute of Molecular Medicine, National Tsing Hua University, Hsinchu 300, Taiwan.|Department of Medical Science, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Linyi
authors.initials: L
authors.lastname: Chen
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Wen-Ching
authors.initials: WC
authors.lastname: Wang
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Wenya
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Chien-Wen
authors.initials: CW
authors.lastname: Chang
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.|Department of Medical Science, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Margaret Dah-Tsyr
authors.initials: MD
authors.lastname: Chang
authors.affiliation: Center of Infectious Diseases and Signal Transduction, National Cheng Kung University, Tainan 701, Taiwan.|Department of Pharmacology, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Meng-Ru
authors.initials: MR
authors.lastname: Shen
authors.affiliation: Center of Infectious Diseases and Signal Transduction, National Cheng Kung University, Tainan 701, Taiwan.|National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Tainan 704, Taiwan.|Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan 710, Taiwan.
authors.forename: Ih-Jen
authors.initials: IJ
authors.lastname: Su
authors.affiliation: Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.|Department of Medical Science, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Lily Hui-Ching
authors.initials: LH
authors.lastname: Wang
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D002462:Cell Membrane / Q000378:metabolism; D043171:Chromosomal Instability; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006509:Hepatitis B / Q000150:complications* / Q000235:genetics / Q000821:virology; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism*; D006515:Hepatitis B virus / Q000472:pathogenicity; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009154:Mutation; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D015220:Calcium Channels; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen; D002118:Calcium
keywords: ER stress; SOCE; aneuploidy; ground-glass hepatocytes; hepatitis B virus