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吴茱萸碱通过不同途径诱导肿瘤细胞死亡:凋亡和坏死
AIM: To study the different death pathways in human cervical cancer HeLa and melanoma A375-S2 cells initiated by evodiamine. METHODS: Viability of evodiamine-induced HeLa and A375-S2 cells was measured by MTT assay. Apoptotic cells with condensed or fragmented nuclei were visualized by Hoechst 33258 staining. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Proportion of cell death through apoptotic and necrotic pathways was determined by LDH activity-based cytotoxicity assays. Cell cycle distribution was observed by flow cytometry. RESULTS: Evodiamine induced HeLa and A375-S2 cell death dose- and time-dependently.Caspase-3 and-8 were activated in apoptosis induced by evodiamine 15 μmol/L. However, over 24- h incubation of A375-S2 cells, evodiamine 15 μmol/L initiated necrosis related to p38 and ERK (extracellular signal-regulated kinases)activities. Evodiamine-induced HeLa cell death was preceded by an accumulation of cells at the G2/M phase of the cell cycle, but there was no significant effect of evodiamine on A375-S2 cell cycle. CONCLUSION: Evodiamine induces caspase-3,8-dependent apoptosis in HeLa cells which is related to G2/M arrest of the cell cycle. On the other hand, in A375-S2 cells, evodiamine initiates caspase-3,8-mediated apoptosis at early stages and the induction of MAPK-mediated necrosis at later stages of cell culture.
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Objective:To explore the proliferation inhibition and apoptosis-inducing effect of evodiamine in human gastric cancer SGC-7901 cells. Methods:After 48 or 24 h exposure to different concentrations of evodiamine, cell proliferation was analyzed using tetrazolium blue (MTT) assay while apoptosis and cell-cycle phase distribution using lfow cytometry. Results:In 0.01~30.00μg/mL range of concentrations, evodiamine inhibited the proliferation of SGC-7901 cells in dose-dependent manner, and the overall mean IC50 was (3.79±0.16)μg/mL;the apoptosis rate was increased from 3.4%to 7.0%, 13.8%and 36.3%at concentrations of 0, 0.5, 1.5 and 30μg/mL of evodiamine, respectively;the percentage of cells accumulated in G2/M phase was increased from 17.26%to 98.92%in the cells treated with evodiamine for 24 h in 0.01~30.00μg/mL range of concentrations. Conclusion:Evodiamine can inhibit the proliferation, induce apoptosis in human gastric cancer cell line SGC-7901 in vitro and arrest the cell cycle at the G2/M phase.