肿瘤细胞膜模型筛选抗肿瘤多肽新方法的建立

Establishment of Tumor Cell Membrane Model and Screening for Specific Anti-tumor Peptides

背景与目的:部分多肽类药物具有靶向选择性抗肿瘤作用,目前针对多肽抗肿瘤的研究都是基于某一或数个肽链的信息,受到现有多肽信息的限制。本研究旨在通过肿瘤细胞膜模型的建立,应用核糖体表达法进行抗肿瘤多肽筛选。方法:选择已知可在肿瘤细胞膜上穿孔的阳性对照多肽Mast21和MastoparanX优化人工合成的肿瘤脂质体细胞膜模型。组建可直接用于体外多肽表达的DNA库,应用核糖体表达法进行特异性抗肿瘤多肽筛选,应用MTT法检测筛选的多肽在体外抗肿瘤效果。结果:应用核糖体体外表达法和脂质体(PE/PS,3∶7)肿瘤细胞膜模型成功地从设计的肽库中筛选出一些肽,体外实验证实部分多肽能抑制非小细胞肺癌细胞株(NCI-H460)生长,而对正常人体纤维细胞CCD-27SK无影响。结论:合适的肿瘤细胞膜模型和核糖体表达法,可从肽库中筛选特异性抗肿瘤多肽,为开发作用机制完全不同的抗肿瘤新药奠定基础。

BACKGROUND &OBJECTIVE:Some polypeptides have targeting selective a nti-tumor effects act on cell membrane, but studies on this field were restrict ed by lack of peptide information. This study was to establish a tumor cell memb rane model, and isolate specific anti-tumor peptides. METHODS: Pore-forming pe ptides Mast21 and MastoparanX were chosen as positive control peptides to optimi ze the artificially synthesized tumor liposome cell membrane model, ribosome dis play system was used to isolate specific anti-tumor peptides from a designed ra ndom DNA library with affinity to membrane model. Anti-tumor effects of the iso lated peptides on non-small cell lung cancer cell line NCI-H460 were analyzed with MTT assay. RESULTS: After 6-round successive selection with improved membr ane model (PE/PS, 3∶7) and ribosome display system, some sequences of peptides were obtained. In vitro experiment confirmed that some of them showed anti-tumo r effects on NCI-H460 cells, 50%inhibitory concentration (IC50) of peptides Se qA3, and SeqB3 of NCI-H460 cells were (22.5±1.2) μmol/L,and (11.3±0.7) μmol /L, while on normal human fibro cell line CCD-27SK were more than 100 μmol/L. CONCLUSION: Specific anti-tumor peptides may be screened out by suitable tumor cell membrane model and ribosome display system, which will help to develop new anti-tumor drugs.