Effects of transfection on inflammatory factor production in LPS-induced HBE cells

Abstract

Transfection is an experimental technique typically used in biological experiments. In this study, we verified whether this technique may cause the release of inflammatory cytokines and affect cell viability. We used lipopolysaccharide (LPS)-induced human bronchial epithelial (16-HBE) cells as a model to evaluate whether cell transfection with Lipofectamine 3000 would affect LPS-induced inflammatory factors in HBE cells. MicroRNA (miRNA) negative control (NC)- and miR-584-mimics were transfected into 16-HBE cell lines. The 584-mimic was used to increase the expression of miR-584, and the NC-mimic was used to add a negative control sequence. After 24h of transfection, the cells were incubated with LPS for another 24h, and the effects on the release of inflammatory cytokines, such as IL-1, IL-6, IL-8, TNF-α, MIP-1, MCP-1α, and cell viability were investigated. The optimal conditions for transfection were evaluated, and cytokine and chemokine mRNA levels were determined. Regardless of the NC- or 584-mimic, the results indicate that the expression of transfected genes in these cells leads to an increase in inflammatory factors and decreased cell viability. Microscope analysis revealed that the number of HBE cells was lower after transfection, and many small vesicles could be observed in the transfected cells, indicating that the insertion of gene vectors may affect the biological activity of HBE cells and experimental results. Results suggest that Lipofectamine 3000 transfected miRNA into HBE cells, providing better transfection rates, however, at the cost of higher toxicity