Inducible overexpression of zebrafishmicroRNA-722suppresses chemotaxis of human neutrophil like cells

Abstract

Neutrophil migration is essential for battling against infections but also drives chronic inflammation. Since primary neutrophils are terminally differentiated and not genetically tractable, leukemia cells such as HL-60 are differentiated into neutrophil-like cells to study mechanisms underlying neutrophil migration. However, constitutive overexpression or inhibition in this cell line does not allow the characterization of the genes that affect the differentiation process. Here we apply the tet-on system to induce the expression of zebrafish miR-722 in differentiated HL-60. Overexpression of miR-722 reduced the mRNA level of genes in the chemotaxis and inflammation pathways, including RAC2. Consistently, cell migration is significantly inhibited upon induced miR-722 overexpression. Together, miR-722 is an evolutionarily conserved suppressor for neutrophil migration and signaling in zebrafish and human. providing the Odyssey imaging system (LI-COR).