The role of metalloproteinase and hypoxia conditions in endometrial cells and embryo implantation

Maryam Mahdavi, Diba Bagheri, Elmira Vanaki, Mehdi Shamsara, Ehsan Hashemi, Saeed Ansari, Mojtaba Dashtizad, Morteza Daliri Joupari


In the process of implantation, metalloproteinase enzymes play a key role in basement membrane degradation and endometrial extracellular matrix. The activity of these enzymes is impeded by binding Tissue Inhibitors of Metalloproteinase (TIMP). The oxygen concentration in the mammalian uterus at the time of implantation is about 2-5%. It is seen that the imposition of hypoxia on cancer cells increases the expression of metalloproteinase enzymes and reduces the expression of metalloproteinase inhibitors, resulting in increased cell invasion. To know the effect of Hypoxia-Inducible Factor (HIF) and other related factors, we decided to evaluate hypoxic conditions on endometrial epithelial cells of the uterus and roll of matrix metalloproteinases (MMPs) on angiogenesis and invasion of the embryo during implantation. In this study, human and mouse endometrial epithelial cells were incubated for 24-48 hours in hypoxic conditions. Subsequently, the expression level of TIMP-1 was measured in mouse and human epithelial cells by Real-Time PCR technique. The cell viability in hypoxic conditions was evaluated by MTT assay. Our results demonstrated that hypoxia reduced the quantitative gene expression of TIMP-1 in the human and mouse endometrial epithelial cells compared to the control group. It can be concluded that applying hypoxic conditions by reducing the TIMP-1 expression and consequently increasing MMP expression, may improve the embryo implantation rate.


Implantation; Embryo; Hypoxia; TIMP-1; MMPs


Koot YE, Macklon NS. Embryo implantation: biology, evaluation, and enhancement. Curr Opin Obstet Gynecol. 2013; 25(4):274-9.

Bischof P, Campana A. A putative role for oncogenes in trophoblast invasion? Hum Reprod. 2000; 15 Suppl 6:51-8.

Gaddy-Kurten D, Hickey GJ, Fey GH, Gauldie J, Richards JS. Hormonal regulation and tissue-specific localization of alpha 2-macroglobulin in rat ovarian follicles and corpora lutea. Endocrinology. 1989; 125(6):2985-95.

Dajee M, Kazansky AV, Raught B, Hocke GM, Fey GH, Richards JS. Prolactin induction of the alpha 2-Macroglobulin gene in rat ovarian granulosa cells: stat 5 activation and binding to the interleukin-6 response element. Mol Endocrinol. 1996; 10(2):171-84.

Nagase H, Woessner JF, Jr. Matrix metalloproteinases. J Biol Chem. 1999; 274(31):21491-4.

Määttä M, Soini Y, Liakka A, Autio-Harmainen H. Localization of MT1-MMP, TIMP-1, TIMP-2, and TIMP-3 messenger RNA in normal, hyperplastic, and neoplastic endometrium. Enhanced expression by endometrial adenocarcinomas is associated with low differentiation. Am J Clin Pathol. 2000; 114(3):402-11.

Williamson RA, Marston FA, Angal S, Koklitis P, Panico M, Morris HR, et al. Disulphide bond assignment in human tissue inhibitor of metalloproteinases (TIMP). Biochem J. 1990; 268(2):267-74.

Staun-Ram E, Shalev E. Human trophoblast function during the implantation process. Reprod Biol Endocrinol. 2005; 3:56.

Kovalenko VL, Kazachkov EL, Voropaeva EE, Kazachkova EA. [The characteristics of the epithelium and endometrial extracellular matrix in miscarriage during early pregnancy associated with chronic endometritis]. Arkh Patol. 2009; 71(5):40-3.

Goffin F, Munaut C, Frankenne F, Perrier D'Hauterive S, Béliard A, Fridman V, et al. Expression pattern of metalloproteinases and tissue inhibitors of matrix-metalloproteinases in cycling human endometrium. Biol Reprod. 2003; 69(3):976-84.

Smith SK. Angiogenesis and implantation. Hum Reprod. 2000; 15 Suppl 6:59-66.

Shifren JL, Tseng JF, Zaloudek CJ, Ryan IP, Meng YG, Ferrara N, et al. Ovarian steroid regulation of vascular endothelial growth factor in the human endometrium: implications for angiogenesis during the menstrual cycle and in the pathogenesis of endometriosis. J Clin Endocrinol Metab. 1996; 81(8):3112-8.

Li S, Cao Y, Zhang J, Tian Y, Zheng X, Duan E. Effects of vascular endothelial growth factor on MMPs during embryo implantation in mice. Chinese Sci Bull. 2002; 47(24):2071-4.

Dong JC, Dong H, Campana A, Bischof P. Matrix metalloproteinases and their specific tissue inhibitors in menstruation. Reproduction. 2002; 123(5):621-31.

Meade ES, Ma YY, Guller S. Role of hypoxia-inducible transcription factors 1alpha and 2alpha in the regulation of plasminogen activator inhibitor-1 expression in a human trophoblast cell line. Placenta. 2007; 28(10):1012-9.

Canning MT, Postovit LM, Clarke SH, Graham CH. Oxygen-mediated regulation of gelatinase and tissue inhibitor of metalloproteinases-1 expression by invasive cells. Exp Cell Res. 2001; 267(1):88-94.

Singh H, Aplin JD. Adhesion molecules in endometrial epithelium: tissue integrity and embryo implantation. J Anat. 2009; 215(1):3-13.

Alvarez-Tejado M, Naranjo-Suarez S, Jiménez C, Carrera AC, Landázuri MO, del Peso L. Hypoxia induces the activation of the phosphatidylinositol 3-kinase/Akt cell survival pathway in PC12 cells: protective role in apoptosis. J Biol Chem. 2001; 276(25):22368-74.

Nie GY, Butt AR, Salamonsen LA, Findlay JK. Hormonal and non-hormonal agents at implantation as targets for contraception. Reprod Fertil Dev. 1997; 9(1):65-76.

Martínez-Hernández MG, Baiza-Gutman LA, Castillo-Trápala A, Armant DR. Regulation of proteinases during mouse peri-implantation development: urokinase-type plasminogen activator expression and cross talk with matrix metalloproteinase 9. Reproduction. 2011; 141(2):227-39.

Menino AR, Jr., O'Claray JL. Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen. J Reprod Fertil. 1986; 77(1):159-67.

Aflalo ED, Sod-Moriah UA, Potashnik G, Har-Vardi I. Expression of plasminogen activators in preimplantation rat embryos developed in vivo and in vitro. Reprod Biol Endocrinol. 2005; 3:7.

Curry TE, Jr., Osteen KG. The matrix metalloproteinase system: changes, regulation, and impact throughout the ovarian and uterine reproductive cycle. Endocr Rev. 2003; 24(4):428-65.

Mastroianni L, Jr., Jones R. Oxygen tension within the rabbit fallopian tube. J Reprod Fertil. 1965; 9:99-102.

Catt JW, Henman M. Toxic effects of oxygen on human embryo development. Hum Reprod. 2000; 15 Suppl 2:199-206.

Fischer B, Bavister BD. Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits. J Reprod Fertil. 1993; 99(2):673-9.

Iyer NV, Kotch LE, Agani F, Leung SW, Laughner E, Wenger RH, et al. Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1 alpha. Genes Dev. 1998; 12(2):149-62.

Lando D, Peet DJ, Whelan DA, Gorman JJ, Whitelaw ML. Asparagine hydroxylation of the HIF transactivation domain a hypoxic switch. Science. 2002; 295(5556):858-61.

Minet E, Ernest I, Michel G, Roland I, Remacle J, Raes M, et al. HIF1A gene transcription is dependent on a core promoter sequence encompassing activating and inhibiting sequences located upstream from the transcription initiation site and cis elements located within the 5'UTR. Biochem Biophys Res Commun. 1999; 261(2):534-40.

Déry MA, Michaud MD, Richard DE. Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol. 2005; 37(3):535-40.

Bos R, van der Groep P, Greijer AE, Shvarts A, Meijer S, Pinedo HM, et al. Levels of hypoxia-inducible factor-1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer. 2003; 97(6):1573-81.

Jung JE, Lee HG, Cho IH, Chung DH, Yoon SH, Yang YM, et al. STAT3 is a potential modulator of HIF-1-mediated VEGF expression in human renal carcinoma cells. Faseb j. 2005; 19(10):1296-8.

Pufe T, Harde V, Petersen W, Goldring MB, Tillmann B, Mentlein R. Vascular endothelial growth factor (VEGF) induces matrix metalloproteinase expression in immortalized chondrocytes. J Pathol. 2004; 202(3):367-74.



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