中华真地鳖抗肿瘤蛋白纯化、鉴定及活性研究

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。

Purification,identification and anticancer activity of a novel protein,from Eupolyphaga sinensis Walker

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 purity 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 fractions.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 significantly 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.