Zhuo Yu1, Yue-Qiu Gao2, Hai Feng3, Ying-Ying Lee4, May S Li3, Yuan Tian4, Minnie Y Y Go4, Dae-Yeul Yu5, Yue-Sun Cheung6, Paul B S Lai7, Jun Yu8, Vincent W S Wong8, Joseph J Y Sung9, Henry L Y Chan8, Alfred S L Cheng10. 1. Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China Department of Liver Disease, Shuguang Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China. 2. Department of Liver Disease, Shuguang Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China. 3. School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 4. Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 5. Disease Model Research Laboratory, Aging Research Center and World Class Institute, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. 6. Department of Surgery, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 7. Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China Department of Surgery, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 8. Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 9. Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China. 10. School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China.
Abstract
BACKGROUND: Androgen receptor (AR) signalling contributes to male predominance in hepatocellular carcinoma (HCC), which is more pronounced in HBV-endemic areas. Cell cycle-related kinase (CCRK) is essential for AR-induced hepatocarcinogenesis but its molecular function in HBV-associated HCC remains obscure. OBJECTIVE: To determine the molecular function of CCRK in HBV-associated HCC. DESIGN: Transcriptional regulation was assessed by chromatin immunoprecipitation, promoter mutation and luciferase reporter assays. Hepatocellular proliferation and tumourigenesis were examined by colony formation, soft agar assays and using HBV X protein (HBx) transgenic mice with low-dose exposure to diethylnitrosamine. Protein expressions were examined in clinical samples and correlated with patient survival by log-rank Mantel-Cox test. RESULTS: Overexpression of CCRK, but not its kinase-defective mutant, activated β-catenin/T cell factor signalling through phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9, led to upregulation of AR transcriptional activity and, subsequently, expression of HBx. The viral transactivator in turn induced CCRK expression through enhanced AR signalling, thus forming a positive regulatory loop. RNA interference silencing of CCRK, which suppressed the CCRK/GSK-3β/β-catenin/AR regulatory loop, significantly suppressed HBx-induced hepatocellular proliferation (p=0.001) and transformation (p<0.001) and remarkably reduced >80% diethylnitrosamine-mediated hepatocarcinogenesis in HBx transgenic mice. Finally, patients with HBV-associated HCC with concordant overexpression of CCRK, GSK-3β phosphorylation at Ser9, active dephosphorylated β-catenin and AR phosphorylation at Ser81 had poorer overall (HR=31.26, p<0.0001) and disease-free (HR=3.60, p<0.01) survival rates. CONCLUSIONS: Our findings highlight the critical role of CCRK in a self-reinforcing circuitry that regulates HBV-associated hepatocarcinogenesis. Further characterisation of this intricate viral-host signalling may provide new prognostic biomarkers and therapeutic targets for HCC treatment. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
BACKGROUND:Androgen receptor (AR) signalling contributes to male predominance in hepatocellular carcinoma (HCC), which is more pronounced in HBV-endemic areas. Cell cycle-related kinase (CCRK) is essential for AR-induced hepatocarcinogenesis but its molecular function in HBV-associated HCC remains obscure. OBJECTIVE: To determine the molecular function of CCRK in HBV-associated HCC. DESIGN: Transcriptional regulation was assessed by chromatin immunoprecipitation, promoter mutation and luciferase reporter assays. Hepatocellular proliferation and tumourigenesis were examined by colony formation, soft agar assays and using HBV X protein (HBx) transgenic mice with low-dose exposure to diethylnitrosamine. Protein expressions were examined in clinical samples and correlated with patient survival by log-rank Mantel-Cox test. RESULTS: Overexpression of CCRK, but not its kinase-defective mutant, activated β-catenin/T cell factor signalling through phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9, led to upregulation of AR transcriptional activity and, subsequently, expression of HBx. The viral transactivator in turn induced CCRK expression through enhanced AR signalling, thus forming a positive regulatory loop. RNA interference silencing of CCRK, which suppressed the CCRK/GSK-3β/β-catenin/AR regulatory loop, significantly suppressed HBx-induced hepatocellular proliferation (p=0.001) and transformation (p<0.001) and remarkably reduced >80% diethylnitrosamine-mediated hepatocarcinogenesis in HBx transgenic mice. Finally, patients with HBV-associated HCC with concordant overexpression of CCRK, GSK-3β phosphorylation at Ser9, active dephosphorylated β-catenin and AR phosphorylation at Ser81 had poorer overall (HR=31.26, p<0.0001) and disease-free (HR=3.60, p<0.01) survival rates. CONCLUSIONS: Our findings highlight the critical role of CCRK in a self-reinforcing circuitry that regulates HBV-associated hepatocarcinogenesis. Further characterisation of this intricate viral-host signalling may provide new prognostic biomarkers and therapeutic targets for HCC treatment. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
Authors: Anees M Dauki; James S Blachly; Esko A Kautto; Sameera Ezzat; Mohamed H Abdel-Rahman; Christopher C Coss Journal: Cancer Res Date: 2019-11-04 Impact factor: 12.701
Authors: Jingying Zhou; Man Liu; Hanyong Sun; Yu Feng; Liangliang Xu; Anthony W H Chan; Joanna H Tong; John Wong; Charing Ching Ning Chong; Paul B S Lai; Hector Kwong-Sang Wang; Shun-Wa Tsang; Tyler Goodwin; Rihe Liu; Leaf Huang; Zhiwei Chen; Joseph Jy Sung; King Lau Chow; Ka Fai To; Alfred Sze-Lok Cheng Journal: Gut Date: 2017-09-22 Impact factor: 23.059