棘刺锚参抗肿瘤因子的纯化及其抗肿瘤作用  

Purification and antitumor effect evaluation of Protankyra bidentata antitumor factor

作　　者：沈先荣[1] 栾洁[1] 蒋定文[1] 陈伟[1] 刘李娜[1] 贾福星[1] 

机构地区：[1]海军医学研究所,上海200433

出　　处：《中国海洋药物》2013年第1期1-6,共6页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(30471972)资助

摘　　要：目的纯化棘刺锚参抗肿瘤因子(Protankyra bidentata Antitumor Factor,PBATF),并探讨其抗肿瘤活性。方法棘刺锚参为原料,采用盐溶液抽提,硫酸胺分级沉淀,DEAE-Sepharose CL-6B离子交换层析,Sephacryl S-300分子筛层析,ZORBAX GF-250HPLC层析,得到PBATF。采用SDS-PAGE电泳和HPLC鉴定其纯度。采用MTT方法测定PBATF对肿瘤细胞生长的影响,克隆形成实验测定对肿瘤细胞集落形成的影响。采用荷瘤小鼠试验测定PBATF对体内肿瘤生长的影响。结果从棘刺锚参纯化获得PBATF,达到较高纯度。PBATF对7901人胃癌细胞、Hela细胞、NCI非小细胞肺癌细胞均具有显著的生长抑制效应,而对929小鼠成纤维细胞和小牛主动脉血管内皮细胞只在高浓度时有抑制作用。PBATF对Hela细胞的集落形成与生长具有明显的抑制作用,试验浓度时对皮肤成纤维细胞的集落形成无明显影响。PBATF对小鼠Lew-is肺癌有明显的疗效,结论获得了高纯度的PBATF,能够显著抑制体内外肿瘤生长。5mg·kg-1剂量具有56.89%的抑瘤率,1mg·kg-1剂量的抑瘤率为52.00%,0.5mg·kg-1剂量的抑瘤率为33.61%。Objective To purify the Protankyra bidentata anti-tumor factor （PBATF） from Protankyra bidentata, and study its antitumor effects. Methods PBATF was extracted from Protankyra bidentata with 10 mmol L-1 PB containing 1 mol L · NaCI, and fractionally precipitated with 40%- 60% （NH4）2SO4. Further purifications were achieved by DEAE-Sepharose CL-6B ion exchange chromatog- raphy, Sephacryl S-300 gel filtration and reverse-phase high performance liquid chromatography. PBATF was evaluated as homogeneous by SDS PAGE and HPLC analyses. MTT and Clone Formation Assay were used to detect the inhibitory effects of PBATF on the growth of tumor cells and the colony formation of cell, respectively. Tumor-bearing mice were used to determine the effect of PBATF on tumor growth in vivo. Results PBATF was purified from Protankyra bidentata, and appeared a single band on a 12% sodium dodecyl sulfate-polyacrylamide gel, and a single peak on Zorbax Eclipse XDB C18 HPLC chromatography. PBATF inhibited the growth of 7901, Hela, and NCI cells significantly. but had no significant effect on 929 cells and bovine artery endothelial cells. PBATF inhibited the colo ny formation of Hela cells, and had no effect on 929 cells. The tumor growth of Lewis lung cancer im planted in C57BL/6 mice was inhibited significantly by PBATF, and the inhibitory rates were 56.89 in 5 mg kg-1 group, 52.00% in 1 mg kg 1 group, 33.61% in 0.5 mg kg 1. Conclusion PBATF purified from Protankyra bidentata has significant inhibitory effect on tumor growth both in vitro and in vivo.

关 键 词：海参 纯化 抗肿瘤 肿瘤细胞 

分 类 号：R965[医药卫生—药理学]