A zebrafish model of intrahepatic cholangiocarcinoma by dual expression of hepatitis B virus X and hepatitis C virus core protein in liver.
The mechanisms that mediate the initiation and development of intrahepatic cholangiocarcinoma (ICC) associated with hepatitis B and C virus (HBV and HCV, respectively) infection remain largely unclear. In this study we conditionally coexpressed hepatitis B virus X (HBx) and hepatitis C virus core (HCP) proteins in zebrafish livers, which caused fibrosis and consequently contributed to ICC formation at the age of 3 months. Suppressing the transgene expression by doxycycline (Dox) treatment resulted in the loss of ICC formation. The biomarker networks of zebrafish ICC identified by transcriptome sequencing and analysis were also frequently involved in the development of human neoplasms. The profiles of potential biomarker genes of zebrafish ICC were similar to those of human cholangiocarcinoma. Our data also showed that the pSmad3L oncogenic pathway was activated in HBx and HCP-induced ICC and included phosphorylation of p38 mitogen-activated proteinbase (MAPK) and p44/42 mitogen-activated protein kinase (ERK1/2), indicating the association with transforming growth factor beta 1 (TGF-β1) signaling pathway in ICC. Bile duct proliferation, fibrosis, and ICC were markedly reduced by knockdown of TGF-β1 by in vivo morpholinos injections.


CONCLUSIONS
These results reveal that TGF-β1 plays an important role in HBx- and HCP-induced ICC development. This in vivo model is a potential approach to study the molecular events of fibrosis and ICC occurring in HBV and HCV infection.
Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan.
Wangta
W
Liu
Jim-Ray
JR
Chen
Chih-Hao
CH
Hsu
Yen-Hsing
YH
Li
Yi-Meng
YM
Chen
Chien-Yuan
CY
Lin
Shin-Jie
SJ
Huang
Zen-Kuei
ZK
Chang
Yen-Chun
YC
Chen
Chi-Hsueh
CH
Lin
Hong-Yi
HY
Gong
Ching-Chun
CC
Lin
Koichi
K
Kawakami
Jen-Leih
JL
Wu
D000818:Animals; D030801:Animals, Genetically Modified; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001650:Bile Duct Neoplasms / Q000235:genetics* / Q000473:pathology; D001653:Bile Ducts, Intrahepatic; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000473:pathology; D055513:Connective Tissue Growth Factor / Q000235:genetics; D019938:Cyclin D1 / Q000235:genetics; D004195:Disease Models, Animal; D004318:Doxycycline / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D016174:Hepacivirus; D020935:MAP Kinase Signaling System; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics; D051899:Smad2 Protein / Q000235:genetics; D051900:Smad3 Protein / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D014758:Viral Core Proteins / Q000235:genetics*; D054334:Viral Regulatory and Accessory Proteins; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics
D000900:Anti-Bacterial Agents; D051899:Smad2 Protein; D051900:Smad3 Protein; D015534:Trans-Activators; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A; D014758:Viral Core Proteins; D054334:Viral Regulatory and Accessory Proteins; D029961:Zebrafish Proteins; C057628:hepatitis B virus X protein; C497786:smad2 protein, zebrafish; D019938:Cyclin D1; D055513:Connective Tissue Growth Factor; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D004318:Doxycycline