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噬菌体展示的角蛋白生长因子模型肽可促进表皮细胞增殖而无致瘤性。

Keratinocyte growth factor phage model peptides can promote epidermal cell proliferation without tumorigenic effect.

机构信息

Institute of Tissue Engineering, Second Hospital, Shandong University, Jinan, Shandong 250033, China.

出版信息

Chin Med J (Engl). 2010 May 5;123(9):1195-200.

Abstract

BACKGROUND

Keratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation.

METHODS

A phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells.

RESULTS

Thirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4).

CONCLUSION

Four phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

摘要

背景

角质细胞生长因子(KGF)对上皮细胞的创伤愈合有显著影响。本研究旨在从噬菌体展示 7 肽文库中分离 KGF 噬菌体模拟肽,以评估其对促进表皮细胞增殖的作用。

方法

用单克隆抗人 KGF 抗体作为靶标筛选噬菌体展示 7 肽文库。采用酶联免疫吸附试验(ELISA)筛选具有良好结合活性的单克隆噬菌体。进行 DNA 测序以找到模型肽的相似性。采用三-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)检测、免疫荧光检测和实时定量 PCR 分析来评价噬菌体模型肽对表皮细胞的影响。

结果

通过 ELISA 检测,从 58 个分离的单克隆噬菌体中,有 33 个噬菌体表现出高结合活性。从 15 个获得的噬菌体模型肽中,有 10 个与 KGF 或表皮生长因子(EGF)相似。MTT 检测数据表明,10 个噬菌体模型肽中有 4 个(No.1-4)能够促进表皮细胞增殖。KGF 对照组和两个噬菌体模型肽组(No.1 和 No.2)的角质细胞生长因子受体(KGFR)mRNA 表达增加。KGF 对照组的 c-Fos mRNA 和 c-Jun mRNA 表达增加,但在四个噬菌体模型肽组(No.1-4)中没有增加。

结论

从噬菌体展示 7 肽文库中分离出的 4 个噬菌体模拟肽能够安全地促进表皮细胞增殖,而无致瘤作用。

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