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Keratinocyte growth factor phage model peptides can promote epidermal cell proliferation without tumorigenic effect
Authors:ZONG Xian-lei  JIANG Du-yin  WANG Ji-chang  LIU Jun-li  LIU Zhen-zhong  CAI Jing-long
Institution:[1]Institute of Tissue Engineering,Second Hospital,Shandong University,Jinan,Shandong 250033,China [2]Department of Plastic and Burn Surgery,Second Hospital,Shandong University,Jinan,Shandong 250033,China [3]Lab of Clinical Molecular Biology,Second Hospital,Shandong University,Jinan,Shandong 250033,China [4]Plastic Surgery Hospital,Chinese Academy of Medical Sciences,Beijing 100144,China
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.
Keywords:phage display peptide library  keratinocyte growth factor  peptide  cellular proliferation  wound healing
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