Effect of human beta-defensin-3 on head and neck cancer cell migration using micro-fabricated cell islands

Kevin Wang,Russell Wang,Harihara Baskaran,Rick Jurevic,Joanne H Wang

doi:10.1186/1758-3284-4-41

Abstract

BackgroundTo examine the effect of the natural antimicrobial peptide human β-defensin-3 (hBD-3), on the migration of a head and neck cancer cell line in vitro using microfabrication and soft-lithographic techniques.MethodsTR146 cancer cells were seeded in Petri dishes with microfabricated wells for cell migration assays. Total 54 cell islands were used of various shape and size and experimental media type. Cell migration assays were analyzed in six group media: Dulbecco’s modified medium (DMEM); DMEM with vascular endothelial growth factor (VEGF); Conditioned media of human embryonic kidney cells (HEK 239) expressing hBD-3 via transfected cloned pcDNA3 as CM/hBD-3; CM/hBD-3 + VEGF; conditioned medium from non-transfected HEK 239 (not expressing hBD-3) as control (CM); and the last group was CM + VEGF. Cell islands were circular or square and varied in size (0.25 mm2, 0.125 mm2, and 0.0625 mm2). Cell islands were imaged at t = 0 h, 3 h, 6 h, and 24 h.ResultsThe results show cancer cell islands that originally were smaller had higher migration indices. There was no difference of MIs between circular and square cell islands. MIs at the end point were significantly different among the groups except between CM and CM-hBD-3+ VEGF.ConclusionsVEGF enhanced cancer cell migration. The combination of DMEM and VEGF showed a synergistic effect on this phenomenon of cancer cell migration. Conditioned medium with hBD-3 suppressed cancer cell migration. hBD-3 suppressed VEGF enhancement of TR146 cancer cell migration.

Highlights

To examine the effect of the natural antimicrobial peptide human β-defensin-3, on the migration of a head and neck cancer cell line in vitro using microfabrication and soft-lithographic techniques
Soft lithography that is derived from photolithography and associated techniques which offers tools for micropatterning that complement and extend conventional fabrication methods for biological research [17]. By using these microfabrication and soft lithography techniques; we examined the effect of a natural antimicrobial peptide, human β-defensin-3, on a human head and neck cell line (TR146) migration in vitro
The medium (CM/Human β-defensin (hBD)-3) was collected and centrifuged at 5,000xg to remove cell debris and again the concentration of human beta defensin-3 (hBD-3) was determine by enzyme-linked immunosorbent assay (ELISA) and solution was diluted to 20 ng/mL. 3 ml of the conditioned medium with 20 ng of hBD-3 was used as the Conditioned media (CM)/hBD-3 solution

Summary

Introduction

To examine the effect of the natural antimicrobial peptide human β-defensin-3 (hBD-3), on the migration of a head and neck cancer cell line in vitro using microfabrication and soft-lithographic techniques. In the head and neck region, two of the most common types of cancer are cancer of the oral cavity and cancer of the oropharynx. More than 35,000 Americans are diagnosed each year with oral or oropharyngeal cancer, which will cause over 8,000 deaths, killing roughly one person per hour. In the 1980s, endogenous antimicrobial peptides (AMPs) were found at high concentrations in the granules of neutrophils and macrophages.

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Conclusion