AIM: Thioacetamide (TAA) has been used in studying liver fibrosis and cirrhosis, however, the mechanisms of TAA-induced apoptosis in liver are still unclear. The hepatic epithelial cell line clone 9 was cultured and treated with TAA to investigate the causes of cell death. METHODS: The cell viability of TAA-induced clone 9 cells was determined using MTT assay. Total cellular GSH in TAA-induced clone 9 cells was measured using a slight modification of the Tietze assay. The activity of caspase 3 in TAA-induced clone 9 cells was monitored by the cleavage of DEVD-p-nitroanaline. TUNEL assay and flow cytometry were applied for the determination of DNA fragmentation and the proportion of apoptosis in TAA-induced clone 9 cells, respectively. The alterations of caspase 3, Bad, Bax and Phospho-P53 contents in TAA-induced clone 9 cells were measured by Western blot. RESULTS: The experimental data indicated that TAA caused rat hepatic epithelial cell line clone 9 cell death in a dose-and time-dependent manner; 60% of the cells died (MTT assay) within 24 h after 100 mg/L TAA was applied. Apoptotic cell percentage (TUNEL assay) and caspase 3 activities were highest after 100 mg/L TAA was added for 8 h. The release of GSH and the elevation in caspase content after TAA treatment resulted in clone 9 cell apoptosis via oxidative stress and a caspase-dependent mechanism. The phospho-p53, Bax and Bad protein expressions in clone 9 cells were increased after TAA treatment. CONCLUSION: These results reveal that TAA activates p53, increases caspase 3, Bax and Bad protein contents, perhaps causing the release of cytochrome c from mitochondria and the disintegration of membranes, leading to apoptosis of cells.
AIM: Thioacetamide (TAA) has been used in studying liver fibrosis and cirrhosis, however, the mechanisms of TAA-induced apoptosis in liver are still unclear. The hepatic epithelial cell line clone 9 was cultured and treated with TAA to investigate the causes of cell death. METHODS: The cell viability of TAA-induced clone 9 cells was determined using MTT assay. Total cellular GSH in TAA-induced clone 9 cells was measured using a slight modification of the Tietze assay. The activity of caspase 3 in TAA-induced clone 9 cells was monitored by the cleavage of DEVD-p-nitroanaline. TUNEL assay and flow cytometry were applied for the determination of DNA fragmentation and the proportion of apoptosis in TAA-induced clone 9 cells, respectively. The alterations of caspase 3, Bad, Bax and Phospho-P53 contents in TAA-induced clone 9 cells were measured by Western blot. RESULTS: The experimental data indicated that TAA caused rat hepatic epithelial cell line clone 9 cell death in a dose-and time-dependent manner; 60% of the cells died (MTT assay) within 24 h after 100 mg/L TAA was applied. Apoptotic cell percentage (TUNEL assay) and caspase 3 activities were highest after 100 mg/L TAA was added for 8 h. The release of GSH and the elevation in caspase content after TAA treatment resulted in clone 9 cell apoptosis via oxidative stress and a caspase-dependent mechanism. The phospho-p53, Bax and Bad protein expressions in clone 9 cells were increased after TAA treatment. CONCLUSION: These results reveal that TAA activates p53, increases caspase 3, Bax and Bad protein contents, perhaps causing the release of cytochrome c from mitochondria and the disintegration of membranes, leading to apoptosis of cells.
Authors: L A Morio; H Chiu; K A Sprowles; P Zhou; D E Heck; M K Gordon; D L Laskin Journal: Toxicol Appl Pharmacol Date: 2001-04-01 Impact factor: 4.219
Authors: M E Caballero; J Berlanga; D Ramirez; P Lopez-Saura; R Gozalez; D N Floyd; T Marchbank; R J Playford Journal: Gut Date: 2001-01 Impact factor: 23.059
Authors: Ruma Mukerjee; Satish L Deshmane; Shongshan Fan; Luis Del Valle; Martyn K White; Kamel Khalili; Shohreh Amini; Bassel E Sawaya Journal: Cell Cycle Date: 2008-09-17 Impact factor: 4.534
Authors: Suzy M Salama; Mahmood A Abdulla; Ahmed S Alrashdi; A Hamid A Hadi Journal: Evid Based Complement Alternat Med Date: 2013-08-07 Impact factor: 2.629
Authors: Suzy M Salama; Ahmed S AlRashdi; Mahmood A Abdulla; Pouya Hassandarvish; Mehmet Bilgen Journal: BMC Complement Altern Med Date: 2013-10-24 Impact factor: 3.659
Authors: Suzy M Salama; Mehmet Bilgen; Ahmed S Al Rashdi; Mahmood A Abdulla Journal: Evid Based Complement Alternat Med Date: 2012-09-04 Impact factor: 2.629
Authors: Paul J Mintz; Kai-Wen Huang; Vikash Reebye; Georgios Nteliopoulos; Hong-Shiee Lai; Pal Sætrom; Noriyuki Kasahara; Steen Jensen; Madhava Pai; Myrtle Y A Gordon; Stephen B Marley; Rosemary Behan; Duncan R Spalding; Abdelali Haoudi; Mohamed M Emara; Joanna Nicholls; John J Rossi; Nagy A Habib Journal: Mol Ther Date: 2013-08-28 Impact factor: 11.454