The key relationship between Sampson’s theory and the presence of mesenchymal stem cells in the menstrual flow (MenSCs), as well as the changes in post-transcriptional regulatory processes as actors in the etiopathogenesis of endometriosis, are poorly understood. No study to date has investigated the imbalance of miRNAs in MenSCs related to the disease. Thus, through literature and in silico analyses, we selected four predicted miRNAs as regulators of EGR1, SNAI1, NR4A1, NR4A2, ID1, LAMC3, and FOSB involved in pathways of apoptosis, angiogenesis, response to steroid hormones, migration, differentiation, and cell proliferation. These genes are frequently overexpressed in the endometriosis condition in our group studies. They were the trigger for the miRNAs search. Therefore, a case–control study was conducted with MenSCs of women with and without endometriosis (ten samples per group). Crossing information obtained from the STRING, PubMed, miRPathDB, miRWalk, and DIANA TOOLS databases, we chose to explore the expression of miR-21-5p, miR-100-5p, miR-143-3p, and miR-200b-3p by RT-qPCR. We found an upregulation of the miR-200b-3p in endometriosis MenSCs (P = 0.0207), with a 7.93-fold change (ratio of geometric means) compared to control. Overexpression of miR-200b has been associated with increased cell proliferation, stemness, and accentuated mesenchymal-epithelial transition process in eutopic endometrium of endometriosis. We believe that dysregulated miR-200b-3p may establish primary changes in the MenSCs, thus favoring tissue implantation at the ectopic site.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
RT-qPCR raw data are available upon request from the corresponding author.
Sourial S, Tempest N, Hapangama DK. Theories on the pathogenesis of endometriosis. Int J Reprod Med. 2014;2014:179515.
Laganà AS, Garzon S, Götte M, et al. The pathogenesis of endometriosis: molecular and cell biology insights. Int J Mol Sci. 2019;20:5615.
Wang Y, Nicholes K, Shih I-M. The origin and pathogenesis of endometriosis. Annu Rev Pathol. 2020;15:71–95.
Sampson JA. Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity. Am J Obstet Gynecol. 1927;14:422–69.
Nikoo S, Ebtekar M, Jeddi-Tehrani M, et al. Menstrual blood-derived stromal stem cells from women with and without endometriosis reveal different phenotypic and functional characteristics. Mol Hum Reprod. 2014;20:905–18.
Sasson IE, Taylor HS. Stem cells and the pathogenesis of endometriosis. Ann N Y Acad Sci. 2008;1127:106–15.
Cousins FL, ODF, Gargett CE. Endometrial stem/progenitor cells and their role in the pathogenesis of endometriosis. Best Pract Res: Clin Obstet Gynaecol. 2018;50:27–38.
Liu Y, Zhang Z, Yang F, et al. The role of endometrial stem cells in the pathogenesis of endometriosis and their application to its early diagnosis. Biol Reprod. 2020;102:1153–9.
Poli-Neto OB, Meola J, Rosa-e-Silva JC, et al. Transcriptome meta-analysis reveals differences of immune profile between eutopic endometrium from stage I-II and III-IV endometriosis independently of hormonal milieu. Sci Rep. 2020;10:313–20.
Poli-Neto OB, Carlos D, Favaretto A, et al. Eutopic endometrium from women with endometriosis and chlamydial endometritis share immunological cell types and DNA repair imbalance: a transcriptome meta-analytical perspective. J Reprod Immunol. 2021;145:103307.
Wren JD, Wu Y, Guo S-W. A system-wide analysis of differentially expressed genes in ectopic and eutopic endometrium. Hum Reprod. 2007;22:2093–102.
Fassbender A, Simsa P, Kyama CM, et al. TRIzol treatment of secretory phase endometrium allows combined proteomic and mRNA microarray analysis of the same sample in women with and without endometriosis. Reprod Biol Endocrinol. 2010;8:1–6.
Fassbender A, Verbeeck N, Börnigen D, et al. Combined mRNA microarray and proteomic analysis of eutopic endometrium of women with and without endometriosis. Hum Reprod. 2012;27:2020–9.
Pan Q, Luo X, Toloubeydokhti T, et al. The expression profile of micro-RNA in endometrium and endometriosis and the influence of ovarian steroids on their expression. Mol Hum Reprod. 2007;13:797–806.
Ohlsson Teague EMC, Van der Hoek KH, Van der Hoek MB, et al. MicroRNA-regulated pathways associated with endometriosis. Mol Endocrinol. 2009;23:265–75.
Chegini N. Uterine microRNA signature and consequence of their dysregulation in uterine disorders. Anim Reprod. 2010;7:117–28.
Filigheddu N, Gregnanin I, Porporato PE, et al. Differential expression of micrornas between eutopic and ectopic endometrium in ovarian endometriosis. J Biomed Biotechnol. 2010;2010:369549.
Hawkins SM, Creighton CJ, Han DY, et al. Functional MicroRNA Involved in Endometriosis. Mol Endocrinol. 2011;25:821–32.
Santamaria X, Taylor H. MicroRNA and gynecological reproductive diseases. Fertil Steril. 2014;101:1545–51.
Braza-Boïls A, Salloum-Asfar S, Marí-Alexandre J, et al. Peritoneal fluid modifies the microRNA expression profile in endometrial and endometriotic cells from women with endometriosis. Hum Reprod. 2015;30:2292–302.
Saare M, Rekker K, Laisk-Podar T, et al. Challenges in endometriosis miRNA studies — from tissue heterogeneity to disease specific miRNAs. Biochim Biophys Acta. 2017;1863:2282–92.
Agrawal S, Tapmeier TT, Rahmioglu N, et al. The miRNA mirage: how close are we to finding a non-invasive diagnostic biomarker in endometriosis? A systematic review. Int J Mol Sci. 2018;19:599.
Mathieu J, Ruohola-Baker H. Regulation of stem cell populations by microRNAs. Adv Exp Med Biol. 2013;786:329–51.
ASRM. Revised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertility and Sterility 1997; 67: 817–821.
Santamaria X, Massasa EE, Taylor HS. Migration of cells from experimental endometriosis to the uterine endometrium. Endocrinology. 2012;153:5566–74.
Cheng Y, Li L, Wang D, et al. Characteristics of human endometrium-derived mesenchymal stem cells and their tropism to endometriosis. Stem Cells Int. 2017;2017:4794827.
Meng X, Ichim TE, Zhong J, et al. Endometrial regenerative cells: a novel stem cell population. J Transl Med. 2007;5:1–10.
Zucherato VS, Penariol LBC, Silva LECM, et al. Identification of suitable reference genes for mesenchymal stem cells from menstrual blood of women with endometriosis. Sci Rep. 2021;11:5422.
Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7.
Musina RA, Belyavski AV, Tarusova OV, et al. Endometrial mesenchymal stem cells isolated from the menstrual blood. Bull Exp Biol Med. 2008;145:539–43.
Szklarczyk D, Gable AL, Lyon D, et al. STRING v11: Protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47:D607–13.
Kehl T, Kern F, Backes C, et al. miRPathDB 2.0: a novel release of the miRNA pathway dictionary database. Nucleic Acids Res. 2020;48:D142–7.
Sticht C, La Torre CD, Parveen A, et al. miRWalk: an online resource for prediction of microRNA binding sites. PLOS ONE. 2018;13:e0206239.
Karagkouni D, Paraskevopoulou MD, Chatzopoulos S, et al. DIANA-TarBase v8: a decade-long collection of experimentally supported miRNA–gene interactions. Nucleic Acids Res. 2018;46:D239–45.
Thermo Fisher Scientific. TaqMan ® Advanced miRNA assays single-tube assays for use with: TaqMan ® Advanced miRNA cDNA Synthesis Kit Catalog Number A25576. user guide 2016; 1–31.
Thermo Fisher Scientific. Application note - amplification efficiency of TaqMan gene expression assays. White Paper 2006; 1–8.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25:402–8.
Du X, Yuan Q, Qu Y, et al. Endometrial mesenchymal stem cells isolated from menstrual blood by adherence. Stem Cells Int. 2016;2016:3573846.
Bozorgmehr M, Gurung S, Darzi S, et al. Endometrial and menstrual blood mesenchymal stem/stromal cells: biological properties and clinical application. Front Cell Dev Biol. 2020;8:497.
Chapron C, Marcellin L, Borghese B, et al. Rethinking mechanisms, diagnosis and management of endometriosis. Nat Rev Endocrinol. 2019;15:666–82.
Ramón LA, Braza-Boïls A, Gilabert-Estellés J, et al. microRNAs expression in endometriosis and their relation to angiogenic factors. Hum Reprod. 2011;26:1082–90.
Yang RQ, Teng H, Xu XH, et al. Microarray analysis of microRNA deregulation and angiogenesis-related proteins in endometriosis. Genetics and Molecular Research 2016; 15: gmr.15027826.
Abdel-Rasheed M, Nour Eldeen G, Mahmoud M, et al. microRNA expression analysis in endometriotic serum treated mesenchymal stem cells. EXCLI J. 2017;16:852–67.
Haikalis ME, Wessels JM, Leyland NA, et al. MicroRNA expression pattern differs depending on endometriosis lesion type. Biol Reprod. 2018;98:623–33.
Abe W, Nasu K, Nakada C, et al. miR-196b targets c-myc and Bcl-2 expression, inhibits proliferation and induces apoptosis in endometriotic stromal cells. Hum Reprod. 2013;28:750–61.
Wright KR, Mitchell B, Santanam N. Redox regulation of microRNAs in endometriosis-associated pain. Redox Biol. 2017;12:956–66.
Zheng B, Xue X, Zhao Y, et al. The differential expression of microRNA-143,145 in endometriosis. Iran J Reprod Med. 2014;12:555–60.
Braza-Boïls A, Marí-Alexandre J, Gilabert J, et al. MicroRNA expression profile in endometriosis: its relation to angiogenesis and fibrinolytic factors. Hum Reprod. 2014;29:978–88.
Saare M, Rekker K, Laisk-Podar T, et al. High-throughput sequencing approach uncovers the miRNome of peritoneal endometriotic lesions and adjacent healthy tissues. PLOS ONE. 2014;9:e112630.
Shi X-Y, Gu L, Chen J, et al. Downregulation of miR-183 inhibits apoptosis and enhances the invasive potential of endometrial stromal cells in endometriosis. Int J Mol Med. 2014;33:59–67.
Logan PC, Yango P, Tran ND. Endometrial stromal and epithelial cells exhibit unique aberrant molecular defects in patients with endometriosis. Reprod Sci. 2017;25:140–59.
Mashayekhi P, Noruzinia M, Zeinali S, et al. Endometriotic mesenchymal stem cells epigenetic pathogenesis: deregulation of miR-200b, miR-145, and let7b in a functional imbalanced epigenetic disease. Cell J. 2019;21:179–85.
Warren LA, Shih A, Renteira SM, et al. Analysis of menstrual effluent: diagnostic potential for endometriosis. Mol Med. 2018;24:1–12.
Chen L, Qu J, Xiang C. The multi-functional roles of menstrual blood-derived stem cells in regenerative medicine. Stem Cell Res Ther. 2019;10:1–10.
Panir K, Schjenken JE, Robertson SA, et al. Non-coding RNAs in endometriosis: a narrative review. Hum Reprod Update. 2018;24:497–515.
Altuvia Y, Landgraf P, Lithwick G, et al. Clustering and conservation patterns of human microRNAs. Nucleic Acids Res. 2005;33:2697–706.
Hu W, Xie Q, Xu Y, et al. Integrated bioinformatics analysis reveals function and regulatory network of miR-200b-3p in endometriosis. Biomed Res Int. 2020;2020:3962953.
Humphries B, Yang C. The microRNA-200 family: small molecules with novel roles in cancer development, progression and therapy. Oncotarget. 2015;6:6472–98.
Jolly MK, Ware KE, Gilja S, et al. EMT and MET: necessary or permissive for metastasis? Mol Oncol. 2017;11:755–69.
Yang Y-M, Yang W-X. Epithelial-to-mesenchymal transition in the development of endometriosis. Oncotarget. 2017;8:41679–89.
Eggers JC, Martino V, Reinbold R, et al. microRNA miR-200b affects proliferation, invasiveness and stemness of endometriotic cells by targeting ZEB1, ZEB2 and KLF4. Reprod Biomed Online. 2016;32:434–45.
Dongre A, Weinberg RA. New insights into the mechanisms of epithelial–mesenchymal transition and implications for cancer. Nat Rev Mol Cell Biol. 2019;20:69–84.
Wellner U, Schubert J, Burk UC, et al. The EMT-activator ZEB1 promotes tumorigenicity by repressing stemness-inhibiting microRNAs. Nat Cell Biol. 2009;11:1487–95.
Park S-M, Gaur AB, Lengyel E, et al. The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev. 2008;22:894–907.
Ohlsson Teague EMC, Print CG, Hull ML. The role of microRNAs in endometriosis and associated reproductive conditions. Hum Reprod Update. 2010;16:142–65.
Barragan F, Irwin JC, Balayan S, et al. Human endometrial fibroblasts derived from mesenchymal progenitors inherit progesterone resistance and acquire an inflammatory phenotype in the endometrial niche in endometriosis. Biol Reprod. 2016;94:118.
The authors thank Océlia de Vasconcelos for her support in collecting the samples. In addition, we are especially grateful to the women who voluntarily agreed to participate in this work.
This study was financially supported by the Sao Paulo Research Foundation (FAPESP 2013/22431–3), National Institute of Hormones and Women’s Health (Hormona)-CNPq (INCT-CNPq 465482/2014–7), and CAPES Higher Education Improvement Coordination (Scholarship).
The Research Ethics Committee of the University Hospital of the Ribeirao Preto Medical School approved this case–control study (HCRP 3644/2019). The MenSC used in this work were collected from November 2014 to December 2016 following the ethics guidelines established by the Declaration of Helsinki (HCRP 15227/2012). They have been transferred to a biorepository of the Human Reproduction Section at the Department of Gynecology and Obstetrics of the Ribeirao Preto Medical School (HCRP 3644/2019).
Consent to Participate
All participants have provided written informed consent (HCRP. 3644/2019, approval date 06/04/2019).
Consent for Publication
All authors have read and approved the manuscript publication.
Conflict of Interest
The authors declare no competing interests.
About this article
Cite this article
de Oliveira, R.Z., de Oliveira Buono, F., Cressoni, A.C.L. et al. Overexpression of miR-200b-3p in Menstrual Blood-Derived Mesenchymal Stem Cells from Endometriosis Women. Reprod. Sci. 29, 734–742 (2022). https://doi.org/10.1007/s43032-022-00860-y