A synthetic peptide inhibits human ovarian cancer cell motility

Abstract

Phage display libraries have been commonly used to identify recognition sites and antigen-antibody binding systems in tumor cells since the 1990s. In the previous study, we isolated a 12-mer binding peptide SVSVGMKPSPRP (SV-peptide) while isolating human ovarian cancer cells (SK-OV-3) using a random phage display library. Therefore, we tested the specific binding and anticancer properties of the SV-peptide with respect to cell viability and motility. The peptide-binding assay showed the immunofluorescent staining intensity of tumor cells (A2780, A549 and SK-OV-3) > control cells (HUVEC). Monolayer scratch healing and Transwell transmigration assays showed that the SV-peptide inhibited the migration of all three cancer cell types but not HUVEC. The mechanism involves a suppression of cytoskeletal F-actin filament arrangement into filopodia and lamellipodia through downregulation of Rac1 but not RhoA. The FAK-PI3K signaling pathway was also affected by the peptide. Western blot analysis revealed a downregulation of FAK, but not of PI3K, after 24 h incubation of SK-OV-3 cells with the SV-peptide. Metalloproteinase-2 (MMP-2) was also downregulated by the SV-peptide. Finally, the viability of cancer cell lines (SK-OV-3 and A549) decreased in the presence of the SV-peptide after 48 h. Transmission electron microscopy of the SV-peptide-exposed cancer cultures showed extensive vacuolization, swollen mitochondria, and rough endoplasmic reticulum. Thus, the SV-peptide has some certain application prospects in the tumor therapy.

Inhibiting Cell Viability and Motility by Layer-by-Layer Assembly and Biomineralization

“…Before the fabrication of the LbL films on the cell surface, we evaluated the cytotoxicity of each solution by the CCK-8 counting kit assay as described previously. 10 The anionic polymer (PSS) showed over 75% viability (Figure 2C), although the cationic polymer solution PDDA showed a greater cytotoxicity (<50% viability) (Figures 2A,B and S8). PDA and ε-PL showed a high cell viability (>80%) even with a cationic charge (Figure 2D,E).…”

“…Before the fabrication of the LbL films on the cell surface, we evaluated the cytotoxicity of each solution by the CCK-8 counting kit assay as described previously. 10 …”

“…In other words, the LbL films should possess certain cellular compatibility. Before the fabrication of the LbL films on the cell surface, we evaluated the cytotoxicity of each solution by the CCK-8 counting kit assay as described previously …”

“…10.1021/acsomega.0c00846.Physiochemical properties of polymers used in this study; chemical structures of the cationic and anionic polymers used in this study; SEM images of the HeLa cells coating with PDDA/PSS films after 24 h of incubation; SEM images of the HeLa cells coating with PDDA/PSS films and calcium phosphate shells after 24 h of incubation; fluorescence microscopic images of PEs films (PDDA/PSS) or mineral shells prepared on the SKOV-3 cells after 24 h incubation; fluorescence microscopic images of PEs films (PDDA/PSS) or mineral shells prepared on the HeLa cells after 24 h incubation; effects of PDDA/PSS films or mineral shells on the migration of cell lines by the scratch wound healing assay; effects of various PE films or mineral shells on the migration of MDA-MB-231 cell lines by the scratch wound healing assay; morphology of HeLa cells before and after coating with PDDA/PSS films and CaCO 3 shells for 1, 3, and 5 days at 37 °C; immunofluorescence analyses of the effects of various PE films or mineral shells on the expression and distribution of Rho A after 24 h coculture; immunofluorescence analyses of the effects of various PE films or mineral shells on the expression and distribution of Cdc 42 after 24 h coculture; concentration of MMP-9 of SKOV-3 cells coated with PDA/GE and CaCO 3 ; and concentration of Rac1 of SKOV-3 cells coated with PDA/GE and CaCO 3 (PDF)Department of Biomedical Engineering, Research Center for Nano-Biomaterials & Regenerative Medicine, College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan 030024, PR China; orcid.org/0000-0002-7927-122X; Phone: +86-351-6014477; Email: weiyan@ tyut.edu.cn; Fax: +86-351-6011816 Di Huang − Department of Biomedical Engineering, Research Center for Nano-Biomaterials & Regenerative Medicine, College of Biomedical Engineering, Taiyuan University of Technology,…”