藜麦活性蛋白的制备及抗结肠癌效应评价

Preparation of Quinoa Active Protein and Effect Evaluation of Its Anti-colon Cancer

本研究以藜麦全谷为原料,采用丙酮-硫酸铵沉淀法经Q柱进一步分离纯化,结合MTT法确定了具有抗结肠癌活性的藜麦蛋白(WQP),并在细胞水平上探究WQP的抗结肠癌活性。采用MTT试验表明WQP能够显著抑制结肠癌DLD1和HCT-8细胞的增殖,并具有质量浓度梯度依赖性,而对人正常结肠上皮细胞FHC无影响;细胞克隆形成试验及细胞形态学试验进一步证明了WQP具有显著的抗结肠癌作用;流式细胞技术检测结果表明WQP能够诱导结肠癌细胞凋亡。此外,细胞划痕试验表明WQP通过抑制DLD1、HCT-8细胞的增殖,阻滞了结肠癌细胞迁移。本研究为将WQP进一步开发成为天然抗结肠癌特医功能食品及保健品提供了理论依据,同时为藜麦蛋白的深度利用奠定了基础。

In this study,quinoa was used as raw material,and the protein component with anti-colon cancer activity was screened by MTT method after further separation and purification by acetone-ammonium sulfate precipitation method through Q column.The anti-colon cancer activity of WQP was further investigated by cell model.The results of cell proliferation assay showed that WQP significantly inhibited the proliferation of colon cancer DLD1 and HCT-8 cells in a concentration gradient dependence,but had no effect on human normal colon epithelial cells FHC.Cell clone formation assay and cell morphology assay further proved that WQP had significant anti-colon cancer effect.Flow cytometry showed that WQP could induce apoptosis of colon cancer cells.In addition,cell scratch assay showed that WQP significantly restrained the migration of DLD1 and HCT-8 cells.Therefore,this study provides a theoretical basis for the development of WQP as a natural anti-colon cancer drug ingredient,and at the same time greatly improves the utilization rate of quinoa protein.