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目的:研究沙苑子黄酮(flavonoids of Astragalws complanatus,FAC)抑制人乳腺癌细胞MCF-7增殖、诱导其凋亡以及FAC对核转录因子κB(nuclear factor-κB,NF-κB)、c-Myc、p53表达的影响。方法:以不同浓度FAC(0.0625、0.125、0.25、0.5mg/ml)处理MCF-7细胞12、24、48、72h后,MTT法测定细胞增殖情况;用流式细胞分析术检测FAC诱导凋亡作用;用荧光免疫法、Western blot法检测凋亡相关蛋白的表达。结果:MTT法显示,不同浓度FAC作用MCF-7细胞24、48、72h后对细胞增殖具有明显抑制作用,与阴性对照组相比差异具有显著性(P<0.05);抑瘤率与FAC剂量、作用时间呈依赖性。流式细胞分析显示,FAC能明显诱导MCF-7细胞凋亡,0.0625、0.125、0.25、0.5mg/ml FAC作用细胞36h后,凋亡率分别为6.5%、18.3%、49.6%和57.9%。荧光免疫法及Western blot法显示,FAC作用后MCF-7中c-Myc与NF-κB表达明显减弱;而p53表达显著增强,并向细胞核内聚集。结论:FAC对MCF-7具有抑制增殖和诱导凋亡作用;其作用可能与其下调NF-кB、c-Myc表达、上调p53表达有关。
OBJECTIVE: To study the effects of flavonoids of Astragalus complanatus (FAC) on the proliferation and apoptosis of human breast cancer cell line MCF-7 and the effect of FAC on the expression of nuclear factor-κB, c-Myc, Effect of p53 expression. Methods: The proliferation of MCF-7 cells treated with FAC (0.0625,0.125,0.25,0.5mg / ml) for 12,24,48,72h were determined by MTT assay. Flow cytometry was used to detect apoptosis induced by FAC The expression of apoptosis related protein was detected by fluorescence immunoassay and Western blot. Results: MTT assay showed that MCF-7 cells treated with different concentrations of FAC had significant inhibitory effect on cell proliferation at 24, 48 and 72 hours (P <0.05), and the inhibition rates were similar to those of FAC , The role of time was dependent. Flow cytometry analysis showed that FAC could induce apoptosis of MCF-7 cells significantly. After treated with 0.0625,0.125,0.25,0.5mg / ml FAC for 36h, the apoptotic rates were 6.5%, 18.3%, 49.6% and 57.9%, respectively. Fluorescent immunofluorescence and Western blot showed that the expression of c-Myc and NF-κB in MCF-7 was significantly decreased after FAC treatment, while the expression of p53 was significantly increased and aggregated to the nucleus. CONCLUSION: FAC can inhibit proliferation and induce apoptosis of MCF-7 cells, which may be related to down-regulation of NF-кB and c-Myc expression and up-regulation of p53 expression.