Doxycycline reduces MMP-2 activity and inhibits invasion of 12Z epithelial endometriotic cells as well as MMP-2 and -9 activity in primary endometriotic stromal cells in vitro

Eleftherios P. Samartzis,Patrick Imesch,Daniel Fink,Manuel Stucki

doi:10.1186/s12958-019-0481-z

Eleftherios P. Samartzis, Patrick Imesch + Show 2 more

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https://doi.org/10.1186/s12958-019-0481-z

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Abstract

BackgroundMatrix metalloproteinases (MMPs), especially the gelatinases MMP-2 and MMP-9, play a crucial role in the pathogenesis of endometriosis by enabling invasion. Doxycycline is a well-tolerated antibiotic and a potent MMP-inhibitor in subantimicrobial doses.MethodsGelatin zymography and activity assays were used to detect latent and active MMP-2 and -9 in cell culture supernatants of immortalized epithelial (12Z) and two isolates of primary endometriotic stromal cells treated with doxycycline. The invasiveness of 12Z endometriotic cells treated with doxycycline was assessed in matrigel-coated invasion chambers. The effect on latent and active MMP-2 expression of the combination of progesterone and doxycycline was tested in 12Z.ResultsDoxycycline significantly reduced the MMP-2 activity and pro-MMP-2 expression in 12Z and the MMP-2 and -9 activity as well as expression of pro-MMP-2 and -9 in primary endometriotic stromal cells. The percentage of 12Z cells invading through a matrigel-coated membrane was reduced to 65 and 22% of the control after treatment with doxycycline at doses of 1 μg/ml and 10 μg/ml, respectively. Furthermore, a combination of progesterone and doxycycline showed an additive effect in low doses on the reduction of MMP-2 activity and pro-MMP2 expression in 12Z endometriotic cells.ConclusionsIn conclusion, the MMP-inhibiting features of subantimicrobial-dose doxycycline may be further evaluated as a well-tolerable additional therapeutic approach, e.g. in combination with progestins such as dienogest, in patients with infiltrative endometriosis with insufficient response to current medical treatment options.

Highlights

Matrix metalloproteinases (MMPs), especially the gelatinases MMP-2 and MMP-9, play a crucial role in the pathogenesis of endometriosis by enabling invasion
Matrix metalloproteinases (MMPs), especially members of the group of gelatinases (MMP-2 and MMP-9), play a crucial role in the development of endometriosis, since MMP-9 has been shown to be increased in eutopic and ectopic endometrial tissue from women with endometriosis and higher levels of MMP-2, − 9, and − 14 mRNA have been found in endometriotic cells when compared to normal endometrium [5,6,7]
We observed no significant cell mortality up to doses of 20 μg/ml doxycycline, whereas 40 μg/ml doxycycline led to a reduced number of viable cells (65% of the control, Fig. 1b). This means that the reduced levels of MMP-2 up to 20 μg/ml were not due to reduced cell numbers or increased mortality of endometriotic cells

Summary

Introduction

Matrix metalloproteinases (MMPs), especially the gelatinases MMP-2 and MMP-9, play a crucial role in the pathogenesis of endometriosis by enabling invasion. One of the most important pathogenic characteristics of the proliferation of endometriosis, especially in the deep-infiltrating form, is the invasion of endometriotic cells through the basilar membrane of the peritoneal mesothelium into the extracellular matrix [1]. Matrix metalloproteinases (MMPs), especially members of the group of gelatinases (MMP-2 and MMP-9), play a crucial role in the development of endometriosis, since MMP-9 has been shown to be increased in eutopic and ectopic endometrial tissue from women with endometriosis and higher levels of MMP-2, − 9, and − 14 mRNA have been found in endometriotic cells when compared to normal endometrium [5,6,7]. The roles and interactions of different MMPs in endometriosis are complex and not yet fully understood [14]

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