[Aminopeptidase inhibitor Bestatin induces HL-60 cell apoptosis through activating caspase 3].
Min-Kai Lin, Jianli He, Zhen Cai, Wei Qian
Abstract
Min-Kai Lin, Jianli He, Zhen Cai, Wei Qian
Abstract
OBJECTIVE: To study the variation and significance of caspase 3 activity in the process of amino-peptidase inhibitor--bestatin (BS) inducing human leukemic cell apoptosis. METHODS: Cell apoptosis was evaluated by light microscopy, TUNEL labeling and flow cytometry (FCM). Caspase 3 activity was detected by colorimetry. The mitochondrial transmembrane potentials (DeltaPsi(m)) were detected by Rhodamine123 staining. RESULTS: The apoptotic morphology, apoptotic peak on FCM and positive Annexin V(FITC) on cell membrane showed that BS could induce HL-60 cell apoptosis in a dose- and time-dependent manner. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The apoptosis induced by BS was inhibited by AC-DEVD-CHO. The DeltaPsi(m) of cells treated with BS declined. CONCLUSION: BS induces apoptosis of human acute leukemic cells through activation of caspase 3.
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OBJECTIVE: To study the variation and significance of caspase 3 activity in the process of amino-peptidase inhibitor--bestatin (BS) inducing human leukemic cell apoptosis. METHODS: Cell apoptosis was evaluated by light microscopy, TUNEL labeling and flow cytometry (FCM). Caspase 3 activity was detected by colorimetry. The mitochondrial transmembrane potentials (DeltaPsi(m)) were detected by Rhodamine123 staining. RESULTS: The apoptotic morphology, apoptotic peak on FCM and positive Annexin V(FITC) on cell membrane showed that BS could induce HL-60 cell apoptosis in a dose- and time-dependent manner. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The apoptosis induced by BS was inhibited by AC-DEVD-CHO. The DeltaPsi(m) of cells treated with BS declined. CONCLUSION: BS induces apoptosis of human acute leukemic cells through activation of caspase 3.
Key concepts: Apoptosis, Annexin, TUNEL assay, Flow cytometry, Molecular biology, Cell biology, Caspase 3, Caspase