骆驼蓬种子中一种具抗肿瘤活性蛋白的分离纯化及鉴定  

Purification and Identification of a Novel Protein with Antitumor Activity from Peganum harmala Seeds

作　　者：马晓瑾[1] 吴婷[1] 罗晶晶[1] 王燕[1] 李阳[1] 刘东亮[1] 孙素荣[1] 

机构地区：[1]新疆大学生命科学与技术学院、新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《天然产物研究与开发》2012年第8期1020-1025,共6页Natural Product Research and Development

基　　金：新疆自治区科技支疆项目(2009911219);新疆高校科研计划重点项目(XJEDU2008I01);新疆生物资源基因工程重点实验室开放基金(XJDX0201-2009-03);新疆大学"天山学者"科研启动项目

摘　　要：骆驼蓬种子经浸提、硫酸铵沉淀、CM阳离子交换层析和Superdex 75凝胶过滤层析分离纯化得到一种具有抗肿瘤细胞增殖活性的蛋白(命名为PhLTP),经Tricine-SDS-PAGE检测为单一蛋白条带,高效液相色谱检测其表观分子量为14.8 kDa左右,表明PhLTP是由两条相同的亚基组成的蛋白。采用Edman降解法对该纯化蛋白的N-末端进行氨基酸测序,其N-末端序列与其他植物非特异性脂转移蛋白相似。对PhLTP抗肿瘤活性进行研究,结果表明其对HeLa、Eca-109、MGC-9和BEL-7404细胞都有增殖抑制活性,其中对HeLa细胞增殖的抑制作用较好,并具有浓度和时间依赖性,其IC50为45μg/mL。通过Hoechst33258染色观察细胞形态,发现PhLTP能诱导HeLa细胞发生凋亡。A novel protein with anti-proliferative activity, namaly PhLTP, was isolated from Peganum harmala seeds by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on CM-Sepharose and gel-filtration on Superdex 75. The apparent molecular mass of the purified Peganum harmala protein was 14.8 kDa, with comprising of homodimer. It has similar N-terminal amino acid sequences with non-specific lipid transfer proteins （nsLTPs） of the other plants using NCBI Blast searching for short. PhLTP showed anti-prollferative activity towards some well established cancer cell lines, e. g. Eca-109, MGC-9, BEL-740, and HeLa. The inhibitory effect on the HeLa cell via- bility was dependent on both concentration and time, and the hemi-inhibitory concentration was 45 μg,/mL after 48 hour. PhLTP was also found that this protein induced apoptosis in HeLa cells as evidenced by cell morphological analysis.

关 键 词：骆驼蓬 脂转移蛋白 分离纯化 抗肿瘤 诱导凋亡 

分 类 号：R284.2[医药卫生—中药学]