关键词:
apoptosis
migration
multiplication
neotuberostemonine
pancreatic cancer
摘要:
Objective To explore the effects and mechanisms of neotuberostemonine on the apoptosis of human pancreatic cancer cells MiaPaCa-2 and PANC-1. Methods MiaPaCa-2 and PANC-1 cells were treated with NTS at concentrations of 0, 20, 40, 60, 80, and 100 μg·mL−1 for 24, 48, and 72 h, respectively. Cell viability was detected by the MTT assay. The control group (0 μg·mL−1), the low, medium, high-concentration group of NTS (20, 40, 60 μg·mL−1), were set up. The effect of NTS on the proliferation ability of MiaPaCa-2 and PANC-1 cells was detected by the plate cloning method. The cell migration ability was detected by the scratch healing assay. Apoptosis was observed by Hoechst 33258 staining. The apoptosis rate and mitochondrial membrane potential were detected by flow cytometry. The expression levels of apoptosis-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), activated Caspase-3 (cleaved-Caspase-3), and Caspase-9 were detected by Western blotting. Results Compared with the control group, different concentrations of NTS significantly reduced the proliferation activity (P < 0.01), colony formation rate (P < 0.05 or 0.01), scratch healing rate (P < 0.05 or 0.01), and mitochondrial membrane potential (P < 0.01) of MiaPaCa-2 and PANC-1 cells. In addition, NTS increased the apoptosis level of MiaPaCa-2 and PANC-1 cells (P < 0.01). At the protein level, NTS significantly upregulated the expression levels of apoptosis-related factors cleaved-Caspase-3, Bax, and Caspase-9 proteins, while the expression level of the apoptosis factor Bcl-2 protein was significantly downregulated (P < 0.05, 0.01). Conclusion NTS can effectively inhibit the proliferation and migration of human pancreatic cancer cells MiaPaCa-2 and PANC-1 and promote their apoptosis. The mechanism may be related to the downregulation of the anti-apoptotic protein Bcl-2 and the upregulation of the pro-apoptotic proteins Bax, cleaved-Caspase-3, and Caspase-9. ©