Feng Xiaozhou, Liu Juanjuan, Fan Shuai, Liu Fan, Li Yadong, Jin Yuanyuan, Bai Liping, Yang Zhaoyong
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People's Republic of China.
Sci Rep. 2016 Apr 14;6:24467. doi: 10.1038/srep24467.
An anticancer bioactive peptide (ACBP), goat peroxiredoxin-5 (gPRDX5), was identified from goat-spleen extract after immunizing the goat with gastric cancer-cell lysate. Its amino acid sequence was determined by employing 2D nano-LC-ESI-LTQ-Orbitrap MS/MS combined with Mascot database search in the goat subset of the Uniprot database. The recombinant gPRDX5 protein was acquired by heterogeneous expression in Escherichia coli. Subsequently, the anti-cancer bioactivity of the peptide was measured by several kinds of tumor cells. The results indicated that the gPRDX5 was a good anti-cancer candidate, especially for killing B16 cells. However, the peptide was found to be unstable without modification with pharmaceutical excipients, which would be a hurdle for future medicinal application. In order to overcome this problem and find an effective way to evaluate the gPRDX5, nanoparticle formation, which has been widely used in drug delivery because of its steadiness in application, less side-effects and enhancement of drug accumulation in target issues, was used here to address the issues. In this work, the gPRDX5 was dispersed into nanoparticles before delivered to B16 cells. By the nanotechnological method, the gPRDX5 was stabilized by a fast and accurate procedure, which suggests a promising way for screening the peptide for further possible medicinal applications.
一种抗癌生物活性肽(ACBP),即山羊过氧化物酶体增殖物激活受体5(gPRDX5),在用胃癌细胞裂解物免疫山羊后,从山羊脾脏提取物中鉴定出来。通过二维纳米液相色谱-电喷雾电离-线性离子阱-轨道阱质谱联用技术(2D nano-LC-ESI-LTQ-Orbitrap MS/MS)结合在通用蛋白质数据库(Uniprot)山羊子集中的 Mascot 数据库搜索,确定了其氨基酸序列。通过在大肠杆菌中的异源表达获得了重组 gPRDX5 蛋白。随后,用几种肿瘤细胞测定了该肽的抗癌生物活性。结果表明,gPRDX5 是一种良好的抗癌候选物,尤其对杀死 B16 细胞有效。然而,发现该肽在未经药用辅料修饰时不稳定,这将成为其未来药物应用的障碍。为了克服这个问题并找到评估 gPRDX5 的有效方法,由于纳米颗粒在应用中具有稳定性、副作用小以及能增强药物在靶组织中的积累等优点,已广泛用于药物递送,因此这里使用纳米颗粒形成来解决这些问题。在这项工作中,将 gPRDX5 分散到纳米颗粒中后再递送至 B16 细胞。通过纳米技术方法,gPRDX5 以快速准确的程序得以稳定,这为筛选该肽以用于进一步可能的药物应用提示了一条有前景的途径。