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采用丙酮沉淀、硫酸铵沉淀、Q阴离子交换柱、SP阳离子交换柱、蛋白热变性法对小米米糠蛋白进行逐步分离纯化,并结合细胞增殖抑制实验(MTT法),筛选纯化出小米米糠中抗肿瘤活性蛋白(FMB)。体外抗肿瘤活性实验表明:不同质量浓度(0.025、0.05、0.075、0.1μg/μL)的FMB均可抑制人结肠癌细胞株DLD1和人宫颈癌细胞株HeLa的增殖,并且具有明显的剂量依赖性;而对正常人肝细胞HL-7702无明显抑制作用。倒置显微镜下观察细胞形态特征发现:经FMB处理48h后,DLD1和HeLa细胞出现细胞核明显皱缩、部分细胞漂浮死亡等凋亡特征;而对正常肝细胞株HL-7702无明显影响。
The rice bran protein was separated and purified by acetone precipitation, ammonium sulfate precipitation, Q anion exchange column, SP cation exchange column and protein thermal denaturation method. The anti-tumor activity of millet bran was detected by MTT assay Active protein (FMB). In vitro antitumor activity experiments showed that FMB with different concentrations (0.025,0.05,0.075,0.1μg / μL) could inhibit the proliferation of human colon cancer cell line DLD1 and human cervical cancer cell line HeLa, and had obvious dose-dependent ; While the normal human hepatocyte HL-7702 no significant inhibitory effect. The morphological characteristics of the cells were observed under an inverted microscope. The results showed that after 48h treatment with FMB, DLD1 and HeLa cells showed obvious shrinkage of the nucleus, some of the cells floating and death, but no obvious effect on the normal liver cell line HL-7702.