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目的分离纯化梅花鹿鹿角脱盘总肽,以小鼠乳腺癌细胞系MA782为体外研究对象,初步研究鹿角脱盘总肽的抗肿瘤活性。方法采用酸提醇沉、真空旋转蒸发及冷冻干燥提取鹿角脱盘总肽,再经Sephadex G-25凝胶过滤层析除盐,BCA法测定总肽的蛋白质浓度,SDS-PAGE电泳分析总肽中的蛋白质,MTT法和流式细胞仪检测鹿角脱盘总肽对小鼠乳腺癌细胞系MA782增殖的影响和细胞周期的变化。结果分离纯化出鹿角脱盘总肽粗提物,其蛋白质含量为91.800%;SDS-PAGE分析显示,鹿角脱盘总肽中主要含有11 kDa,18 kDa,40 kDa,64 kDa和80 kDa 5种多肽,总肽对小鼠乳腺癌细胞的增殖具有明显抑制作用,细胞周期各期细胞比例变化明显,当总肽浓度为30 mg/L时,作用72 h后细胞抑制率最高为69.200%,G0/G1期细胞百分比为81.621%,S期细胞百分比为8.983%。结论鹿角脱盘总肽对乳腺癌细胞具有抑制作用,为深入研究鹿角脱盘的抗肿瘤活性提供了实验依据。
OBJECTIVE To isolate and purify total antitumor peptide of antler deer antler, and to investigate the antitumor activity of the total antler of antler offspring by using the mouse breast cancer cell line MA782 as in vitro study. Methods The total antapoptotic peptide of antler was extracted by acid-alcohol precipitation, vacuum rotary evaporation and freeze-drying. The total peptide was separated by Sephadex G-25 gel filtration chromatography. The total protein concentration was determined by BCA method. The total peptide was analyzed by SDS- The protein, MTT assay and flow cytometry were used to detect the effect of antler off-total peptide on the proliferation of mouse breast cancer cell line MA782 and the cell cycle. Results The crude extract of antler off-chip total peptide was separated and purified with a protein content of 91.800%. SDS-PAGE analysis showed that the total peptide of antler offspring contained 11 kDa, 18 kDa, 40 kDa, 64 kDa and 80 kDa The peptide and the total peptide had a significant inhibitory effect on the proliferation of mouse breast cancer cells, and the proportion of cells in the cell cycle changed significantly. When the total peptide concentration was 30 mg / L, the highest cell inhibition rate was 69.200% / G1 phase cell percentage was 81.621%, S-phase cell percentage was 8.983%. Conclusion The antler off-chip total peptide has an inhibitory effect on breast cancer cells, providing an experimental basis for further study antitumor activity of antler.