Tumor necrosis factor α up-regulates endometrial milk fat globule-epidermal growth factor 8 protein production via nuclear factor κB activation, resulting in cell migration of epithelial cells

Abstract

Objective: To explore the role of tumor necrosis factor (TNF) α, an early embryonic product, on endometrial epithelial cell migration and endometrial milk fat globule-epidermal growth factor 8 protein (MFG-E8) production.

Design: In vitro study.

Setting: Academic center.

Intervention(s): Ishikawa cells, used as surrogates for human epithelial cells, were treated with and without TNF-α.

Main outcome measure(s): Effect of TNF-α on intracellular MFG-E8 protein was evaluated with the use of ELISA, Western blot, and subcellular fractionation. Specific inhibitors were used to study TNF-α mechanism of action. Effect of TNF-α on cell migration was studied with the use of a wound healing assay and reorganization of E-cadherin.

Result(s): TNF-α induced: 1) significant up-regulation of MFG-E8 intracellular protein, which was attenuated by pretreatment with a specific inhibitor of nuclear factor κB; 2) increased transcription of MFG-E8 and other proinflammatory factors, such as interleukins 6 and 8, which were suppressed by cotreatment with hCG; and 3) significant cell migration with E-cadherin remodeling, changes associated with subcellular MFG-E8 relocalization.

Conclusion(s): TNF-α up-regulates endometrial epithelial cell migration and MFG-E8 production, which are critical steps required for the endometrial changes during menstrual cycle as well as during embryonic attachment and invasion.

Keywords: Human endometrium; MFG-E8; TNF-α; cell migration.