小米米糠中抗癌细胞增殖活性蛋白的分离纯化

doi:10.7506/spkx1002-6630-201309060

食品科学 ›› 2013, Vol. 34 ›› Issue (9): 296-300.doi: 10.7506/spkx1002-6630-201309060

小米米糠中抗癌细胞增殖活性蛋白的分离纯化

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,*

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

收稿日期:2012-06-06 修回日期:2013-03-19 出版日期:2013-05-15 发布日期:2013-05-07
通讯作者: 单树花 E-mail:kuaile_yezi@163.com
基金资助:
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)

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

SHAN Shu-hua1,2，WU Hai-li1，LI Zong-wei1，YUAN Lin-jie1，ZHAO Wen-bin1，LIU Qing-wu1，GUO Mao-lin1，LI Zhuo-yu1,*

1. Key Laboratory of Chemical Biology and Molecular Engineering, Ministry of Education, Institute of Biotechnology,
Shanxi University, Taiyuan 030006, China；2. Department of Biology, Taiyuan Normal University, Taiyuan 030031, China

Received:2012-06-06 Revised:2013-03-19 Online:2013-05-15 Published:2013-05-07
Contact: shan shuhua E-mail:kuaile_yezi@163.com

摘要/Abstract

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。

关键词: 小米米糠蛋白, 分离纯化, 抗癌细胞, 增殖活性

Abstract:

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.

Key words: foxtail millet bran protein (FMB), isolation and purification, cancer cells, proliferation inhibitory activity

中图分类号:

Q946

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,*. 小米米糠中抗癌细胞增殖活性蛋白的分离纯化[J]. 食品科学, 2013, 34(9): 296-300.

SHAN Shu-hua1,2，WU Hai-li1，LI Zong-wei1，YUAN Lin-jie1，ZHAO Wen-bin1，LIU Qing-wu1，GUO Mao-lin1，LI Zhuo-yu1,*. Isolation and Purification of a Protein from Foxtail Millet Bran and Its Anti-proliferation Activities against Cancer Cells[J]. FOOD SCIENCE, 2013, 34(9): 296-300.

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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

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