海洋岩虫抗菌肽筛选及抗癌活性的初步研究

Screening of antimicrobial peptide from marine clamworm Marphysa sanguinea and preliminary research on its antitumor effect

将岩虫匀浆液经反相浓缩、凝胶过滤和亲和柱层析获得活性组分,在体外用MTS/PMS(氮兰四唑盐/吩嗪硫酸甲酯)方法筛选抗茵肽,再分别用MTS/PMS方法和ELISA-AFP(甲胎蛋白)方法检测其对人肝癌细胞株HepG2增殖、AFP分泌及对正常大鼠成骨细胞株MC3T3—E1增殖的影响。结果表明,纯化的岩虫抗茵肽为组成型碱性蛋白(MW 8.1 kDa,pI8.6),不同浓度抗茵肽对肝癌细胞增殖和AFP的分泌均有抑制作用,抑制作用大小与抗菌肽浓度呈正相关性;对正常成骨细胞未见明显的抑制和杀伤作用。

Active fractions were obtained from the c(?)amworm homogenate by reverse-phase concentration, gel filtration and affinity column chromatography. The MTS/PMS assay was used in the screening of antimicrobial peptide in vitro. The MTS/PMS assay and ELISAAFP method were combined to evaluate the effects of antimicrobial peptide on the proliferation and α-fetoprotein secretion of HepG2 cells as well as on the proliferation of normal mouse osteoblast MC3T3-E1. The results showed that the purified antimicrobial peptide from clamworm Murphysa sanguinea was a constitutive basic protein with MW 8.1 kDa and pI 8.6, which showed inhibition effects on the proliferation and α-fetoprotein secretion of HepG2 cells at different levels. These effects have a positive correlation with the concentrations of antimicrobial peptide. No obvious inhibition and damage effects were observed on the normal osteoblast.