大鲵皮肤黏液糖蛋白的提取纯化及抗肺癌活性研究  

Extraction and purification of giant salamander skin mucous glycoprotein and study its anti-cancer activity of lung cancer

作　　者：徐伟良[1] 陈德经[2] 刘宇[3] 魏泓[3] 刘青 

机构地区：[1]陕西理工学院生物科学与工程学院,陕西汉中723000 [2]陕西理工学院陕西省资源生物重点实验室,陕西汉中723000 [3]第三军医大学基础部实验动物学教研室,重庆400038

出　　处：《中国生化药物杂志》2015年第8期44-47,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：陕西省中小企业科技创新项目(2014CXZJ36)

摘　　要：目的研究大鲵皮肤黏液糖蛋白提取纯化工艺以及体外对人肺癌细胞A549活性的影响。方法利用碱提取和DEAE-52离子交换层析与Sephadex G-100凝胶层析分离纯化大鲵黏液糖蛋白;并采用MTT比色法体外检测大鲵黏液糖蛋白对人肺癌细胞A549的抑制率。结果黏液糖蛋白中的总糖含量为4.23%,经SDS-PAGE电泳检测,糖蛋白的分子量在30 k Da左右,为单一纯品。随糖蛋白纯品浓度从1、10、20、40μg/m L逐渐增加,糖蛋白对A549细胞的抑制率逐渐增大;当糖蛋白浓度为40μg/m L,作用24 h时,对A549细胞的抑制率可达85.66%,作用48 h时,可达92.32%。与阳性对照紫杉醇相比,大鲵皮肤黏液糖蛋白对人肺癌细胞A549有显著的抑制作用。结论大鲵黏液糖蛋白对人肺癌细胞具有明显的抑制作用,可为抗肺癌药物的开发提供理论依据。Objective To study the appearance of skin mucous glycoprotein in vitro on the activity of human lung cancer cells A549. Methods Used alkali extraction and DEAE-52 ion exchange chromatography and Sephadex G-100 gel chromatography purification salamander mucous glycoprotein; Salamander mucous glycoprotein inhibition of human lung cancer cells A549 was detected by MTT colorimetric method in vitro.ResuIts It showed that the total sugar content in the appearance of mucus was 4.23%, the relatively pure glycoprotein component, by SDS protein electrophoresis tests, it contained a single glycoprotein component, its molecular weight was about 30 kDa.With glycoprotein pure concentration increased from 1,10, 20,40μg/mL, the glycoprotein inhibition rate of A549 cells increased; when the glycoprotein concentration was 40 μg/mL, for 24 h action, the inhibition rate of A549 cells was up to 85.66 %, while the role of 48 h, the inhibition rate of A549 cells was up to 92.32%.Inhibition effect of mucus glycoprotein on A549 cell compared with positive control drug, had significant difference.ConcIusion Salamander mucus glycoproteins of human lung cancer cells has obvious inhibitory effect, can provide theoretical basis for the development of lung cancer drug resistance.

关 键 词：大鲵 皮肤黏液 糖蛋白 纯化 MTT法 抗肺癌 

分 类 号：TQ460.1[化学工程—制药化工]