少棘蜈蚣抗菌肽scolopin 2的克隆、表达与抗癌机制研究  

Cloning,Expression and Anticancer Mechanism of Scolopin 2,an Antimicrobial Peptide from Centipede Venoms(Scolopendra subspinipes mutilans

作　　者：侯欢欢[1] 卢佳[1] 章纬菁 黄芳[1] 任文华[1] 

机构地区：[1]南京师范大学生命科学学院,南京210046

出　　处：《生物技术通报》2016年第9期172-178,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31370401);江苏省高校优势学科建设工程项目(PAPD)

摘　　要：旨在研究AMP-scolopin 2的抗菌活性和抗癌机理。利用SUMO融合技术对AMP-scolopin 2进行了体外表达;采用抑菌圈、MTT、流式细胞术和Western blotting等技术分析了其抗菌活性、溶血活性和对肿瘤细胞的促凋亡作用。结果表明,成功表达并纯化了AMP-scolopin 2;AMP-scolopin 2对白血病细胞(K562)和肝细胞癌(Hep G2)具有抑制增殖和促凋亡作用,但对红细胞和正常细胞HEK293的影响很小;AMP-scolopin 2(40μmol/L)可以诱导K562(21.4%)和Hep G2(18.5%)细胞凋亡;此外,AMP-scolopin 2下调了与线粒体相关的凋亡蛋白pro-caspase-3、pro-caspase-9和pro-PARP的表达,且呈浓度依赖性。AMP-scolopin2可以通过线粒体caspase依赖性途径促进肿瘤细胞凋亡。This work is to explore the antimicrobial activity and anticancer mechanism of AMP-scolopin 2. The small ubiquitin-related modifier （ SUMO ） fusion technology was applied in the expression of AMP-scdopin 2 in vitro. The techndogies of inhibition zone assay, MTT, cytotoxicity assay, flow cytometry and Western blotting were utilized to analyze its antimicrobial and hemolytic activity as well as apoptotic effects on cancer cells. As results, AMP-scolopin 2 was successfully expressed and purified. The AMP-scolopin 2 inhibited the proliferation of K562 leukemia cells and HepG2 hepatocellular carcinoma, while it had only a minimal effect on erythrocytes and normal HEK293 cells. AMP- scolopin 2 （ 40 ~tmol/L ） induced the apoptosis of 21.4% K562 leukemia ceils and 18.5% HepG2 hepatocellular carcinoma respectively. In addition, AMP-scolopin 2 down-regulated the expression level of apoptosis proteins （pro-caspase-3, -9 and pro-PARP ） that were related to mitochondria in a dose-dependent mode. In summary, the present study indicates that AMP-scolopin 2 promotes the apoptosis of tumor cells in vitro through mitochondrial caspase-dependent pathway.

关 键 词：蜈蚣 抗菌肽scolopin 2 SUMO 抗菌活性 细胞凋亡 

分 类 号：R285[医药卫生—中药学] R284[医药卫生—中医学]