Hepatitis B virus X induces inflammation and cancer in mice liver through dysregulation of cytoskeletal remodeling and lipid metabolism

// Zhongwei Xu 1, 7 , Linghui Zhai 1 , Tailong Yi 1, 4 , Huiying Gao 1 , Fengxu Fan 1, 4 , Yanchang Li 1 , Youliang Wang 2 , Ning Li 1 , Xiaohua Xing 1 , Na Su 1 , Feilin Wu 1 , Lei Chang 1 , Xiuli Chen 6 , Erhei Dai 6 , Chao Zhao 5 , Xiao Yang 2 , Chunping Cui 1 , Ping Xu 1, 3, 4 1 State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences Beijing, Institute of Radiation Medicine, Beijing, 102206, P.R. China 2 Beijing Institute of Bioengineering, Beijing, 100071, P. R. China 3 Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences, Wuhan, 430072, P. R. China 4 Anhui Medical University, Hefei, 230032, China 5 Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, and Research Center on Aging and Medicine, Fudan University, Shanghai, 200032, China 6 The Fifth Hospital of Shijiazhuang City, Shijiazhuang, 050021, China 7 Central Laboratory, Logistics University of Chinese People’s Armed Police Force, Tianjin, 300309, China Correspondence to: Ping Xu, email: xuping@mail.ncpsb.org Keywords: HBx, SILAM, CDC42, CFL1, ADFP Received: March 10, 2016      Accepted: September 13, 2016      Published: September 30, 2016 ABSTRACT Hepatitis B virus X protein (HBx) participates in the occurrence and development processes of hepatocellular carcinoma (HCC) as a multifunctional regulation factor. However, the underlying molecular mechanism remains obscure. Here, we describe the use of p21 HBx/+ mouse and SILAM (Stable Isotope Labeling in Mammals) strategy to define the pathological mechanisms for the occurrence and development of HBx induced liver cancer. We systematically compared a series of proteome samples from regular mice, 12- and 24-month old p21 HBx/+ mice representing the inflammation and HCC stages of liver disease respectively and their nontransgenic wild-type (WT) littermates. Totally we identified 22 and 97 differentially expressed proteins out of a total of 2473 quantified proteins. Bioinformatics analysis suggested that the lipid metabolism and CDC42-induced cytoskeleton remodeling pathways were strongly activated by the HBx transgene. Interestingly, the protein-protein interaction MS study revealed that HBx directly interacted with multiple proteins in these two pathways. The same effect of up-regulation of cytoskeleton and lipid metabolism related proteins, including CDC42, CFL1, PPARγ and ADFP, was also observed in the Huh-7 cells transfected with HBx. More importantly, CFL1 and ADFP were specifically accumulated in HBV-associated HCC (HBV-HCC) patient samples, and their expression levels were positively correlated with the severity of HBV-related liver disease. These results provide evidence that HBx induces the dysregulation of cytoskeleton remodeling and lipid metabolism and leads to the occurrence and development of liver cancer. The CFL1 and ADFP might be served as potential biomarkers for prognosis and diagnosis of HBV-HCC.