Association between MMP1 and MMP9 activities and ICAM1 cleavage induced by tumor necrosis factor in stromal cell cultures from eutopic endometria of women with endometriosis

Abstract

Endometriosis is a benign gynecological pathology in which immune system deregulation may play a role in its initiation and progression. In endometriotic lesions, intercellular adhesion molecule-1 (ICAM1) is released from the cell membrane by proteolytic cleavage of its extracellular domain, a process that coincides with increased expression and proteolytic activity of metalloproteinases such as MMP1 and MMP9. The objective of our study was to investigate the association between MMP1 and MMP9 activities and ICAM1 cleavage mediated by tumor necrosis factor (TNF) in eutopic endometrial stromal cells from women with and without (control) endometriosis during culture. The RNA was evaluated by RT-PCR, and the protein was determined by western blot (ICAM1, MMP1), casein or gelatin zymographies (secreted active MMP1 or MMP9 respectively), ELISA (soluble ICAM1 (sICAM1)), and fluorescence assay (secreted active MMP1). Under basal conditions, proMMP9 dimer and MMP9 were higher in endometriosis cell cultures. In stromal cultures derived from control women and those with endometriosis, TNF augmented the intracellular proMMP1 (1.2-fold in control stromal cells) and ICAM1 (1.4-and 1.9-fold), greatly increased MMP1 and proMMP9 levels, and the sICAM1 concentration (2.3-and 4.3-fold) in their media compared with basal levels. The combination of TNF and MMP9 increased the sICAM1 concentration 14-fold in the endometriosis cell media, whereas GM6001 inhibited the stimulatory effect of TNF in both cell cultures. The deregulation of MMP9, and the TNF participation in the MMP1 and proMMP9 secretions, in the MMP9 expression and in the expression and cleavage of ICAM1 may contribute to the pathophysiology of this disease.

Identification of Intercellular Cell Adhesion Molecule 1 (ICAM-1) as a Hypoglycosylation Marker in Congenital Disorders of Glycosylation Cells

“…ICAM-1 is a transmembrane protein with five extracellular Ig-like domains and nine occupied N-glycosylation sites in CHO cells (17). It is constitutively expressed by epithelial cells and normally expressed at low levels on immune system and endothelial cells, but it is dramatically upregulated in response to inflammatory stimuli (18,19). As shown by SILAC quantitative peptide pairs in Fig.…”

P450Arom induction in isolated control endometrial cells by peritoneal fluid from women with endometriosis

“…The endometrium was washed, minced, and digested as described by Pino et al (27). The glands were obtained as indicated previously (28).…”

“…The glands were obtained as indicated previously (28). Cells of second passages were cultured until subconfluence, incubated for 24 hours in defined medium (27), and treated (24 hours) with PF-E, PF-C, adsorbed PF-E, N6-2 0 -O-dibutyryl cyclic adenosine 3 0 ,5 0 -monophosphate [(Bu) 2-cAMP, 10 À6 mol/L], or indomethacin (10 À7 -10 À5 mol/L, COX inhibitor) added 30 minutes before the PF-E addition.…”

Hexane extract of aged black garlic reduces cell proliferation and attenuates the expression of ICAM-1 and VCAM-1 in TNF-α-activated human endometrial stromal cells

“…14. It has been shown that the abnormal production of TNF-α contributes to the pathophysiology of endometriosis (18).…”