[TRAIL inhibits proliferation and promotes apoptosis of 3AO ovarian cancer cells].
Shaohua Zhang, Kaige Liu, Biping Cheng, Qian Gao, Li Wang, Xiaofeng Yang
Abstract
Shaohua Zhang, Kaige Liu, Biping Cheng, Qian Gao, Li Wang, Xiaofeng Yang
Abstract
OBJECTIVE: To investigate the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the proliferation of ovarian carcinoma 3AO cells in vitro and its molecular mechanism. METHODS: Ovarian carcinoma 3AO cells were incubated with 12.5, 25.0, 50.0 ng/mL TRAIL proteins for 72 hours, respectively. At 24, 48 and 72 hours, the cells were collected to observe the morphological change under an inverted microscope, detect the cell proliferation using methyl thiazolyl tetrazolium (MTT) assay, the apoptosis rate and cell cycle using flow cytometry (FCM), the morphological features of apoptotic cells using TdT-mediated-dUTP nick end labeling (TUNEL) and the expression of caspase-3 protein using Western blotting. RESULTS: The growth of 3AO cells was inhibited by different concentrations of TRAIL (P<0.05). Morphological change of 3AO cells was clearly observed. TRAIL protein at 25 and 50 ng/mL significantly induced 3AO cell apoptosis and cell cycle arrest. With the treatment time went by, the percentage of cells at G1 phase increased and cells at S and G2/M phase decreased. The expression of caspase-3 protein was raised by TRAIL. No significant differences were noted in the apoptosis rate and the expression of caspase-3 protein between the 25 and 50 ng/mL TRAIL groups (P>0.05). CONCLUSION: TRAIL protein could promote the apoptosis of 3AO cells through inhibiting cell growth cycle, blocking DNA synthesis and activating caspase-3.
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OBJECTIVE: To investigate the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the proliferation of ovarian carcinoma 3AO cells in vitro and its molecular mechanism. METHODS: Ovarian carcinoma 3AO cells were incubated with 12.5, 25.0, 50.0 ng/mL TRAIL proteins for 72 hours, respectively. At 24, 48 and 72 hours, the cells were collected to observe the morphological change under an inverted microscope, detect the cell proliferation using methyl thiazolyl tetrazolium (MTT) assay, the apoptosis rate and cell cycle using flow cytometry (FCM), the morphological features of apoptotic cells using TdT-mediated-dUTP nick end labeling (TUNEL) and the expression of caspase-3 protein using Western blotting. RESULTS: The growth of 3AO cells was inhibited by different concentrations of TRAIL (P<0.05). Morphological change of 3AO cells was clearly observed. TRAIL protein at 25 and 50 ng/mL significantly induced 3AO cell apoptosis and cell cycle arrest. With the treatment time went by, the percentage of cells at G1 phase increased and cells at S and G2/M phase decreased. The expression of caspase-3 protein was raised by TRAIL. No significant differences were noted in the apoptosis rate and the expression of caspase-3 protein between the 25 and 50 ng/mL TRAIL groups (P>0.05). CONCLUSION: TRAIL protein could promote the apoptosis of 3AO cells through inhibiting cell growth cycle, blocking DNA synthesis and activating caspase-3.
Key concepts: Apoptosis, Flow cytometry, Cell cycle, Molecular biology, TUNEL assay, Cell growth, Biology, Cell