桑葚花色苷的提取及对人乳腺癌细胞株MDA-MB-453生长的抑制

Inhibitory effect of anthocyanin extract of mulberry fruit on growth of breast cancer MDA-MB-453 cells

目的以超声辅助乙醇浸提法提取桑葚花色苷,观察其对人乳腺癌细胞株MDA-MB-453体内外生长的抑制作用。方法将桑葚冷冻干燥,冷冻干粉于常温下用70%乙醇以1∶15的料液比超声提取1 h,提取液采用AmberliteTMXAD7HP型大孔吸附树脂以1 ml/min的吸附流速和1 ml/min的洗脱流速在pH3.0酸性环境下纯化,得到桑葚花色苷提取物。利用CCK-8法检测不同浓度桑葚花色苷提取物(50、100、150、200 mg/ml)对人乳腺癌细胞MDA-MB-453增殖活力的影响;以24只雌性BALB/c裸鼠建立MDA-MB-453细胞移植瘤模型,随机分为对照组、低剂量组、中剂量组和高剂量组,分别以0、50、100、200 mg/(kg.d)的桑葚花色苷提取物膳食干预,检测其对肿瘤生长的影响。结果获得花色苷含量为(41.8±4.8)%的桑葚花色苷提取物;CCK-8法检测发现桑葚花色苷提取物可显著降低乳腺癌细胞活力,并显示剂量-效应关系[50、100、150、200 mg/ml桑葚花色苷提取物处理组抑制率分别为(14.8±1.8)%、(29.8±2.2)%、(33.0±2.7)%、(44.3±3.8)%];体内实验发现,高剂量组和中剂量组移植瘤体积和质量均显著低于对照组[体积抑制率分别为(55.5±4.8)%、(39.3±3.2)%;质量抑制率分别为(51.6±4.2)%、(38.9±3.1)%],表明桑葚花色苷提取物膳食干预可有效抑制移植瘤的生长。结论桑葚花色苷提取物能够显著抑制人乳腺癌细胞株MDA-MB-453的生长。

Objective To study the extraction of anthocyanins from mulberry fruits and to observe their inhibitory effect on the growth of human breast cancer MDA-MB-453 cells in vitro and in vivo.Methods Freeze-dried mulberry fruit powder was subjected to ultrasonic extraction with 70% ethanol at a material-to-liquid ratio of 1∶15 for 1 h.The extract solution was purified by AmberliteTM XAD7HP resin in pH 3.0 acidic environment,at an adsorption flow rate 1 ml/min and an elution flow rate 1 ml/min,to obtain an anthocyanin-containing extract.The inhibition of the anthocyanin-containing extract on MDA-MB-453 cell viability was detected by CCK-8 assay.Twenty-four female BALB/c nude mice were injected with MDA-MB-453 cells to establish a transplanted tumor model.The mice were then randomized into 4 groups： a control group,a low dose group,a middle dose group and a high dose group and the treated with the anthocyanin-containing extract at 0,50,100,and 200 mg/（kg·d）,respectively.Tumor volume was measured every four days,and tumor weight was measured on the 28th day.Results The average content of anthocyanins in the mulberry fruit extract was 41.8%.CCK-8 assay indicated that the anthocyanin-containing mulberry fruit extract reduced breast cell viability markedly,exhibiting a dose-effect relationship[（14.8±1.8）%,（29.8±2.2）%,（33.0±2.7）%,（44.3±3.8）%,respectively].Tumors in the high dose and middle dose groups were significantly smaller [（55.5±4.8）% and（39.3±3.2）%] and lighter [（51.6±4.2）% and（38.9±3.1）%] than those in the control group,indicating growth inhibition of the transplanted tumors.Conclusion The anthocyanin-containing mulberry fruit extract can significantly inhibit the growth of human breast cancer MDA-MB-453 cells.