槐米中槲皮素提纯、鉴定以及诱导人乳腺癌MCF-7细胞凋亡作用

目的:研究从槐米中槲皮素(Quercetin,Que)提取、鉴定和对人乳腺癌MCF-7细胞生长的抑制以及诱导凋亡作用.方法:采用碱溶解酸沉淀法提纯槐米中槲皮素,并进行红外光谱分析；采用体外培养人乳腺癌MCF-7细胞,用不同浓度槲皮素处理；采用四甲基偶氮唑蓝(MTT)比色法测定细胞生长抑制率；采用细胞凋亡DNA Lad　der和FITC-Annexin V/PI荧光标记流式细胞仪检测,槲皮素作用后MCF-7细胞凋亡情况.结果:不同浓度槲皮素对人乳腺癌MCF 7细胞有生长抑制作用,并呈浓度和时间依赖性(P＜0.05)；10.0 mmol/L槲皮素作用MCF-7细胞2d,DNA电泳出现特征性的凋亡条带；10.0 mmol/L槲皮素处理MCF-7细胞1d、2d和3d,细胞凋亡率分别为(9.2±1.5)％、(30.0±11.8)％和(60.8±10.6)％.结论:用碱溶解酸沉淀法可从槐米中提纯槲皮素,提取的槲皮素对人乳腺癌MCF-7细胞有生长抑制和诱导凋亡作用.

Effects of quercetins extracted and isolated from flos sophorae immaturus and apoptosis in MCF-7 human breast cancer cell in vitro

AIM: To investigate the effects of quercetins extracted and isolated from the flos sophorae immaturus and apoptosis in MCF-7 human breast cancer cell.METHODS: With the alkali-soluble acid precipitation methods,the quercetins were extracted from flos sophorae immaturus and were identificated by Infrared spectrophotometry.Colorimetric MTT assay was used to measure the prolification inhibitions of the quercetin in MCF-7 cells.The apoptosis of treated cells were examined by DNA agarose gel electrophoresis analysis of DNA fragmentation and flow cytometry using fluorescein labeling FITC-Annexin V/PI,respectively.RESULTS:MTT results showed that the different doses of quercetin can inhibit the growth of MCF-7 cells in dose-and time-dependent significantly(P<0.05).The characteristic bands of apoptosis by DNA electrophoresis were found in MCF-7 cells treated by quercetin at 10.0 mmol/L for 2 d.The rates of apoptosis of MCF-7 cells which were treated with quercetin at 10.0 mmol/L for 1 d,2 d and 3 d were(9.2±1.5)%,(30.0±11.8)% and(60.8±10.6)%,respectively.CONCLUSION: The quercetins can be purified from the flos sophorae immaturus with the alkali-soluble acid precipitation methods.The quercetin can inhibit the growth and apoptosis in MCF-7 cells.