Potent cell growth inhibitory effects in hepatitis B virus X protein positive hepatocellular carcinoma cells by the selective cyclooxygenase-2 inhibitor celecoxib

Huahong Xie; Liucun Gao; Na Chai; Jiugang Song; Jun Wang; Zhenshun Song; Caiping Chen; Yanglin Pan; Lina Zhao; Shiren Sun; Kaichun Wu; Mark A Feitelson; Jie Liu; Daiming Fan

doi:10.1002/mc.20455

Potent cell growth inhibitory effects in hepatitis B virus X protein positive hepatocellular carcinoma cells by the selective cyclooxygenase-2 inhibitor celecoxib

Mol Carcinog. 2009 Jan;48(1):56-65. doi: 10.1002/mc.20455.

Authors

Huahong Xie¹, Liucun Gao, Na Chai, Jiugang Song, Jun Wang, Zhenshun Song, Caiping Chen, Yanglin Pan, Lina Zhao, Shiren Sun, Kaichun Wu, Mark A Feitelson, Jie Liu, Daiming Fan

Affiliation

¹ State Key Laboratory of Cancer Biology, Department of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi Province, People's Republic of China.

PMID: 18506760
DOI: 10.1002/mc.20455

Abstract

Hepatitis B virus (HBV) X protein (HBx) and cyclooxygenase-2 (COX-2) are all playing roles in hepatocellular carcinoma (HCC), but the reversing effects of COX-2 inhibitors on the neoplastic features caused by HBx protein is still unclear. To further evaluate the therapeutic potential of celecoxib on HBx mediated transformation, HCC cells transfected with HBx gene were treated with COX-2 selective inhibitor, celecoxib. The amount the main metabolite of COX-2, prostaglandin E2 (PGE2), was determined by using high sensitivity ELISA. Electron microscope and flow cytometry was used to analyze cell apoptosis and cell cycle distribution. RT-PCR and Western blot were used to identify the molecules involved in celecoxib induced cell apoptosis. The results showed that celecoxib inhibited cell growth more significantly and also induced more cell apoptosis in HBx over-expression cells than in control cells. Celecoxib could selectively inhibited COX-2 expression and PGE2 production. Celecoxib also inhibited p(473Ser)Akt, raf and p53 expression, and induced apoptosis by release of cytochrome c and activation of caspase 9, 3, and 6, which were more remarkably in HBx positive cells than in control cells. These results suggest that celecoxib had potent cell growth inhibitory effects on HBx positive HCC cells mainly through inducing more cell apoptosis, and these findings provide a new insight into the anticancer effects of celecoxib against HBx related HCC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apoptosis / drug effects*
Blotting, Western
Carcinoma, Hepatocellular / enzymology
Carcinoma, Hepatocellular / pathology*
Carcinoma, Hepatocellular / virology
Caspase Inhibitors
Caspases / metabolism
Celecoxib
Cell Cycle / drug effects
Cell Proliferation / drug effects
Cyclooxygenase 2 / chemistry
Cyclooxygenase 2 / metabolism
Cyclooxygenase 2 Inhibitors / pharmacology*
Dinoprostone / metabolism
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Fluorescent Antibody Technique
Humans
Immunoblotting
Immunoenzyme Techniques
Immunoprecipitation
Liver Neoplasms / enzymology
Liver Neoplasms / pathology*
Liver Neoplasms / virology
Phosphorylation / drug effects
Proto-Oncogene Proteins c-akt / metabolism
Proto-Oncogene Proteins c-raf / metabolism
Pyrazoles / pharmacology*
RNA, Messenger / genetics
RNA, Messenger / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sulfonamides / pharmacology*
Trans-Activators / physiology*
Transfection
Tumor Cells, Cultured
Tumor Suppressor Protein p53 / metabolism
Viral Regulatory and Accessory Proteins / physiology*

Substances

Caspase Inhibitors
Cyclooxygenase 2 Inhibitors
Pyrazoles
RNA, Messenger
Sulfonamides
TP53 protein, human
Trans-Activators
Tumor Suppressor Protein p53
Viral Regulatory and Accessory Proteins
hepatitis B virus X protein
Cyclooxygenase 2
PTGS2 protein, human
Proto-Oncogene Proteins c-akt
Proto-Oncogene Proteins c-raf
Caspases
Celecoxib
Dinoprostone