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      A novel mouse CD133 binding-peptide screened by phage display inhibits cancer cell motility in vitro.

      Clinical & Experimental Metastasis

      Animals, Antigens, CD, metabolism, Carrier Proteins, pharmacology, Cell Adhesion, drug effects, Cell Line, Tumor, Cell Movement, Cell Proliferation, Fluorescent Antibody Technique, Glycoproteins, Humans, Male, Mice, Mice, Inbred BALB C, Neoplastic Stem Cells, physiology, Peptide Library, Peptides, Proto-Oncogene Proteins c-met, analysis, genetics, STAT3 Transcription Factor

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          Increased expression of CD133 (Prominin-1), an important cancer stem cell-associated marker, has been observed in the cancer stem cells of a variety of human and mouse cancers. However, no natural ligand of CD133 has yet been identified and little is known about its function. In the present study, LS-7 (amino acid sequence: LQNAPRS), a specific binding peptide targeting mouse CD133, was screened and identified for the first time by phage-displayed peptide library technology. The in vitro and in vivo affinity and specificity of LS-7 were determined, and MTT, adhesion, and migration assays were performed to evaluate the effects of LS-7 on the biological behaviors of cancer cells. To determine which signaling pathways are affected by LS-7, HMGB1, S-100A4, CXCR7, uPAR, AMFR, STAT3, and c-Met gene and protein expression were evaluated by RT-PCR and Western blot. Flow cytometry and immunofluorescence assays showed specific, high-affinity binding of the peptide to mCD133 in vitro. Confocal microscopy confirmed the co-localization of LS-7 positive cells and CD133-positive cells. Migration and wound-healing assays showed that LS-7 significantly inhibited the migration of colon and breast cancer cells in a concentration-dependent manner. In vivo experiments also confirmed the high specificity and affinity of LS-7 to mCD133. RT-PCR and Western blot showed that the expressions of only c-Met and STAT3 decreased obviously in colon and breast cancer cells exposed to LS-7. These findings may provide a novel tool for anti-motility and anti-metastasis strategies in cancer research and cancer stem cell therapy.

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