HBx co-localizes with COXIII in HL-7702 cells to upregulate mitochondrial function and ROS generation

Zou,Lai-Yu; Zheng,Bi-Yun; Fang,Xue-Fen; Li,Dan; Huang,Yue-Hong; Chen,Zhi-Xin; Zhou,Lin-Ying; Wang,Xiao-Zhong

doi:10.3892/or.2015.3852

HBx co-localizes with COXIII in HL-7702 cells to upregulate mitochondrial function and ROS generation

Authors:
- Lai-Yu Zou
- Bi-Yun Zheng
- Xue-Fen Fang
- Dan Li
- Yue-Hong Huang
- Zhi-Xin Chen
- Lin-Ying Zhou
- Xiao-Zhong Wang
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Affiliations: Department of Infection, Union Hospital of Fujian Medical University, Gulou, Fuzhou, Fujian 350001, P.R. China, Department of Gastroenterology, Union Hospital of Fujian Medical University, Gulou, Fuzhou, Fujian 350001, P.R. China, Laboratory of Electron Microscopy, Fujian Medical University, Gulou, Fuzhou, Fujian 350001, P.R. China
Published online on: March 13, 2015 https://doi.org/10.3892/or.2015.3852
Pages: 2461-2467

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Abstract

Hepatocellular carcinoma (HCC) is one of the most common malignant diseases, and HBx leads to the development of HBV-associated HCC. Mitochondria are key organelles that regulate apoptosis, cellular energetics and signal transduction pathways, and are the source of HBx-induced reactive oxygen species (ROS). Recent findings have shown that HBx interacts with the inner mitochondrial membrane protein, COXIII, via the yeast two‑hybrid system, mating experiment and coimmunoprecipitation. The aim of the present study was to examine the co-localizaiton of HBx and COXIII in HL-7702 cells and to investigate ensuing alterations of mitochondrial function. An HL-7702 cell line stably expressing the HBx gene by lentivirus vectors was constructed. Confocal microscopy was utilized to assess the interaction between HBx protein and COXIII. Expression of COXIII, activities of cytochrome c oxidase (COX) and the mitochondrial membrane potential, which were functionally relevant to the HBx protein-COXIII interaction, were investigated in cell cultures. Moreover, the intracellular ROS levels were detected by flow cytometry. The results demonstrated that HBx co-localized with the inner mitochondrial protein, COXIII, in HL-7702 cells, causing the upregulation of COXIII protein expression as well as COX activity. However, HBx did not alter the mitochondrial membrane potential and mitochondria exhibited only slight swelling in HL-7702-HBx cells. Moreover, HBx elevated the generation of mitochondrial ROS in HL-7702-HBx cells. The main finding of the present study was that the co-localization of HBx and COXIII leads to upregulation of the mitochondrial function and ROS generation, which are associated with the oncogenesis of HBV-associated HCC.

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