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Overexpression of cyclooxygenase-2 in human HepG2, Bel-7402 and SMMC-7721 hepatoma cell lines and mechanism of cyclooxygenase-2 selective inhibitor celecoxib-induced cell growth inhibition and apoptosis.
- Source :
-
World journal of gastroenterology [World J Gastroenterol] 2005 Oct 28; Vol. 11 (40), pp. 6281-7. - Publication Year :
- 2005
-
Abstract
- Aim: To investigate the cyclooxygenase-2 (COX-2) expression level in human HepG2, Bel-7402 and SMMC-7721 hepatoma cell lines and the molecular mechanism of COX-2 selective inhibitor celecoxib-induced cell growth inhibition and cell apoptosis.<br />Methods: Hepatoma cells were cultured and treated with celecoxib. Cell in situ hybridization (ISH) and immunocytochemistry were used to detect COX-2 mRNA and protein expression. Proliferating cell nuclear antigen and phosphorylated Akt were also detected by immunocytochemistry assay. Cell growth rates were assessed by 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium (MTT) bromide colorimetric assay. Celecoxib-induced cell apoptosis was measured by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and flow cytometry (FCM). The phosphorylated Akt and activated fragments of caspase-9, caspase-3 were examined by Western blotting analysis.<br />Results: Increased COX-2 mRNA and protein expression were detected in all three hepatoma cell lines. Celecoxib could significantly inhibit cell growth and the inhibitory effect was in a dose- and time-dependent manner evidenced by MTT assays and morphological changes. The apoptotic index measured by TUNEL increased correspondingly with the increased concentration of celecoxib and the reaction time. With 50 micromol/L celecoxib treatment for 24 h, the apoptotic index of HepG2, BEL-7402 and SMMC-7721 cells was 25.01+/-3.08%, 26.40+/-3.05%, and 30.60+/-2.89%, respectively. Western blotting analysis showed remarkable activation of caspase-9, caspase-3 and dephosphorylation of Akt (Thr(308)). Immunocytochemistry also showed the reduction of PCNA expression and phosphorylation Akt (Thr(308)) after treatment with celecoxib.<br />Conclusion: COX-2 mRNA and protein overexpression in HepG2, Bel-7402 and SMMC-7721 cell lines correlate with the increased cell growth rate. Celecoxib can inhibit proliferation and induce apoptosis of hepatoma cell strains in a dose- and time-dependent manner.
- Subjects :
- Apoptosis physiology
Caspase 3
Caspase 9
Caspases metabolism
Celecoxib
Cell Line, Tumor
Cell Proliferation drug effects
Cyclooxygenase 2 genetics
Enzyme Activation
Humans
In Situ Hybridization
Proto-Oncogene Proteins c-akt metabolism
RNA, Messenger metabolism
Apoptosis drug effects
Carcinoma, Hepatocellular
Cyclooxygenase 2 metabolism
Cyclooxygenase 2 Inhibitors pharmacology
Liver Neoplasms
Pyrazoles pharmacology
Sulfonamides pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1007-9327
- Volume :
- 11
- Issue :
- 40
- Database :
- MEDLINE
- Journal :
- World journal of gastroenterology
- Publication Type :
- Academic Journal
- Accession number :
- 16419156
- Full Text :
- https://doi.org/10.3748/wjg.v11.i40.6281