小米米糠中抗癌细胞增殖活性蛋白的分离纯化  

Isolation and Purification of a Protein from Foxtail Millet Bran and Its Anti-proliferation Activities against Cancer Cells

作　　者：单树花[1,2] 武海丽[1] 李宗伟[1] 袁琳洁[1] 赵文彬[1] 刘庆午[1] 郭茂林[1] 李卓玉[1] 

机构地区：[1]山西大学生物技术研究所,化学生物学与分子工程教育部重点实验室,山西太原030006 [2]太原师范学院生物系,山西太原030031

出　　处：《食品科学》2013年第9期296-300,共5页Food Science

基　　金：国家自然科学基金项目(31040018;81150026);高等学校博士学科点专项科研基金项目(20111401110011);山西省国际合作项目(2011081058);山西省留学基金项目(201010)

摘　　要：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。FMB,a novel protein with cancer cell proliferation inhibitory activity was purified from foxtail millet bran by using acetone precipitation,ammonium sulfate precipitation,Q-anion exchange column,SP-cation exchange column and protein thermal denaturation.Its anti-proliferation activities against cancer cells were studied by MTT assay.The results showed the FMB with varied concentrations（0.025,0.05,0.075 μg/μL and 0.1 μg/μL） could significantly inhibit the proliferation of human colonic-cancer cell lines（DLD1） and cervical carcinoma cell line（HeLa） in dose-dependent manner.However,under similar conditions,no significant anti-proliferation effect was observed on normal liver cell line（HL-7702）.Inverted microscope observations revealed that the DLD1 and Hela cells displayed classic morphological features of apoptosis,e.g.,obviously shrinking,floating and partial cell death,after being treated with FMB for 48 h.By contrast,the cell line HL-7702 had no obvious changes under similar conditions.

关 键 词：小米米糠蛋白 分离纯化 抗癌细胞 增殖活性 

分 类 号：Q946[生物学—植物学]