摘要
目的分离纯化中华真地鳖(Eupolyphaga sinensis Walker)中抗肿瘤活性成分并研究其活性。方法采用硫酸铵沉淀、超滤、CM-Sepharose FF、DEAE-Sepharose FF及Q-Sepharose HP离子交换、Butyl Sepharose HP疏水色谱以及Superdex 75凝胶过滤色谱等技术分离纯化抗肿瘤组分;SDS-PAGE及MALDI-TOF质谱进行鉴定;采用MTT法考察其抗肿瘤活性。结果分离纯化得到一分子质量约为72 kD的抗肿瘤蛋白(EPS72)。该蛋白对肝癌Bel-7402细胞、肺癌A549细胞等多种人癌细胞株表现出较强的增殖抑制作用,并呈浓度依赖性。结论纯化的蛋白具有潜在的抗肿瘤作用。
Purpose To report the purification and identification of a novel antitumor protein from E. sinensis and then to investigate its antitumor activities in vitro. Methods The antitumor bioactive fraction was isolated and purified from E. sinensis by using ammonium sulfate precipitation, ultrafiltration, CM- Sepharose FF cation exchange, DEAE-Sepharose FF anion exchange, Q-Sepharose HP anion exchange, Butyl Sepharose HP hydrophobic chromatography and Superdex 75 gel filtration chromatography. The puri- ty and molecular weight of the purified fraction were determined by using SDS-PAGE and MALDI-TOF mass spectra analysis. MTT assay was used to analyze the in vitro antitumor activity of the purified frac- tions. Results The antitumor protein was purified by using a line of chromatographic techniques. And this protein,designated EPS72, showed a unique band with a molecular mass of about 72 kD as determined by SDS-PAGE and gave a mass peak of m/z 71. 737 kD as shown by MALDI-TOF mass spectra. EPS72 sig- nificantly inhibited the proliferation of Bel-7402, A549, RKO, Hela and human pancreatic cancer Panc-28 cells in a dose-dependent manner. Conclusion The purified protein has the potential to be utilized as a novel natural anticancer agent in the future.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2012年第5期519-523,共5页
Chinese Journal of Biochemical Pharmaceutics
关键词
中华真地鳖
抗肿瘤蛋白
纯化
鉴定
活性
Eupolyphaga sinensis Walker
protein
purification
identification
anticancer activity
