基于<i>Musca domestica</i> cecropin的抗肿瘤小分子衍生肽筛选及构效关系解析<sup>*</sup>

doi:10.13523/j.cb.2105028

中国生物工程杂志

2021, Vol. 41

Issue (11): 14-22 DOI: 10.13523/j.cb.2105028

研究报告

基于Musca domestica cecropin的抗肿瘤小分子衍生肽筛选及构效关系解析^*

邓蕊^1,^2,³,曾佳利^1,²,卢雪梅^1,^2,^**(

)

1 广东药科大学生命科学与生物制药学院广州 510006
2 广东省生物活性药物研究重点实验室广州 510006
3 广东药科大学药学院广州 510006

Screening and Structure-activity Relationship Analysis of Anti-tumor Derived Peptides Based on Musca domestica cecropin

DENG Rui^1,^2,³,ZENG Jia-li^1,²,LU Xue-mei^1,^2,^**(

)

1 School of Lif Science and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006,China
2 Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances,Guangzhou 510006,China
3 School of Pharmacy,Guangdong Pharmaceutical University, Guangzhou 510006,China

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摘要：

目的:分析家蝇抗菌肽(Musca domestica cecropin,Mdc)的结构,并对其衍生肽的抗肿瘤活性进行筛选,以期找到高效、经济的抗肿瘤候选小分子肽。方法:采用生物信息学相关软件对抗菌肽Mdc的理化性质和二级结构等进行分析;MTT法测定小分子衍生肽对10种不同肿瘤细胞株的抑制作用,并测定Mdc和衍生肽对正常细胞的毒性作用;溶血实验检测Mdc和衍生肽的溶血性。结果:在抗菌肽Mdc结构分析的基础上,设计了5种衍生肽M1-6、M1-7、M1-8、M9-21、M27-39;MTT结果显示5种衍生肽对特定肿瘤细胞株具有不同程度的抑制作用,但对正常细胞无明显细胞毒性;并且5种衍生肽在一定浓度范围内无溶血现象。结论:成功筛选出了具有相应抗肿瘤活性的小分子肽,其中M27-39、M1-8和M1-7有望成为有前途的抗肿瘤候选小分子肽。

关键词： 抗菌肽; 家蝇天蚕素; 抗肿瘤活性筛选; 构效关系

Abstract:

In order to find more effective, simpler, less toxic, and more stable small peptides, Musca domestica cecropin (Mdc), a housefly antimicrobial peptide cloned in our laboratory, was used to design five derived peptides and analyze the specific structure and anticancer activities of them. Based on the analysis results of Mdc, five peptides derived from the conserved sequence of Mdc were redesigned:M1-6,M1-7,M1-8,M9-21 and M27-39. Bioinformatics tools were used to predict the physical and chemical properties and secondary structure. Proliferation effects of the derived peptides on ten different cancer cell lines and the toxic effects of them on normal cells were determined by MTT assay. The safety of the five polypeptides was verified by hemolysis test and normal cytotoxicity test. Five kinds of derived peptides were designed with Mdc as template, and they all had inhibitory effects on different cancer cells. M1-7, M1-8 and M27-39 showed good activity to several kinds of cancer cells and low hemolytic activity to human erythrocytes. Interestingly, M1-7 had significant inhibitory effect on PC-12 cells, M1-8 had effective biological activity on HepG2 cells, and M27-39 had obvious inhibitory effect on HCT116 cells. The small molecular peptides with corresponding anticancer activity were successfully screened, among which M1-7, M1-8 and M27-39 have obvious specific anticancer activities and may become promising anticancer agents.

Key words: Antimicrobial peptide Musca domestica cecropin Anti-tumor activity screening Structure-activity relationship

收稿日期: 2021-05-17 出版日期: 2021-12-01

ZTFLH:

Q816

基金资助: * 国家重点研发计划(2018YFC1603900);国家自然科学基金(32070509);国家自然科学基金(31501894);广东省基础与应用基础研究专项(2018A030313461)

通讯作者: 卢雪梅 E-mail: luxuemei@gdpu.edu.cn

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引用本文:

邓蕊,曾佳利,卢雪梅. 基于Musca domestica cecropin的抗肿瘤小分子衍生肽筛选及构效关系解析^*[J]. 中国生物工程杂志, 2021, 41(11): 14-22.

DENG Rui,ZENG Jia-li,LU Xue-mei. Screening and Structure-activity Relationship Analysis of Anti-tumor Derived Peptides Based on Musca domestica cecropin. China Biotechnology, 2021, 41(11): 14-22.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2105028 或 https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I11/14

表1 Mdc及5种衍生肽的序列

表2 Mdc及5种衍生肽的理化特性分析结果

图1 ExPASY对Mdc的序列分析

图2 PyMol模拟分析Mdc及5种衍生肽的二级结构

图3 螺旋轮图分析Mdc及5种衍生肽的亲疏水性

图4 ExPASY对Mdc的疏水性分析

图5 5种衍生肽对不同肿瘤细胞增殖的影响

图6 Mdc及5种衍生肽对正常细胞Chang Liver、NCM460细胞毒性研究

图7 不同浓度Mdc及5种衍生肽对人源和鼠源红细胞溶血的影响

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