Expression of miR-126 and Crk in endometriosis: miR-126 may affect the progression of endometriosis by regulating Crk expression

Purpose To evaluate the relationship between miR-126 and Crk and discuss the role of miR-126 in the development and progression of endometriosis (EMs).

The expression levels of miR-126 and Crk mRNA were quantified using real time fluorescent quantitative polymerase chain reaction (real time PCR) in ectopic endometrium (ECs) and eutopic endometrium (EUs) in patients with EMs and normal endometrium (ENs) in EMs-free subjects. The expression levels of Crk protein in all samples were evaluated by Western blot.

The expression level of miR-126 was significantly downregulated in ECs versus EUs (p = 5.45E−5) in the experimental group and in EUs versus ENs (p = 0.019). The expression level of Crk mRNA did not distinguish ECs from EUs (p = 0.995) but was overexpressed in EUs versus ENs (p = 0.006). Crk protein was overexpressed in ECs versus EUs (p = 0.002) in the experimental group and in EUs versus ENs (p = 1.13E−6). The expression level of miR-126 had no correlation with Crk mRNA (p = 0.496) but was negatively correlated with Crk protein (p = 3.134E−5). The expression level of miR-126 in EUs and ECs was negatively correlated with American Fertility Society (AFS) stage (p = 0.022, p = 0.025) and AFS score (p = 0.002, p = 0.007). miR-126 expression decreased with the progression of EMs, but the decrease was not significantly different.

miR-126 may play an initial role in the development and progression of EMs. Crk may be regulated by miR-126, and synergism between abnormal expressions may play an important role in the pathogenesis of EMs. Similar content being viewed by others

Eskenazi B, Warner ML (1997) Epidemiology of endometriosis. Obstet Gynecol Clin North Am 24(2):235–258 Garai J, Molnar V, Varga T, Koppan M, Torok A, Bodis J (2006) Endometriosis: harmful survival of an ectopic tissue. Front Biosci 11:595–619 Sampson JA (1927) Metastatic or embolic endometriosis, due to the menstrual dissemination of endometrial tissue into the venous circulation. Am J Pathol 3(2):93–110.43 Blumenkrantz MJ, Gallagher N, Bashore RA, Tenckhoff H (1981) Retrograde menstruation in women undergoing chronic peritoneal dialysis. Obstet Gynecol 57(5):667–670 Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM (1984) Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol 64(2):151–154 Witz CA, Cho S, Centonze VE, Montoya-Rodriguez IA, Schenken RS (2003) Time series analysis of transmesothelial invasion by endometrial stromal and epithelial cells using three-dimensional confocal microscopy. Fertil Steril 79(Suppl 1):770–778 Aghajanpour L, Mashayekhi F, Rajaei F (2011) Intercellular adhesion molecule-1 (ICAM-1) gene polymorphism and endometriosis in northern Iran. Arch Gynecol Obstet 283(5):1035–1039 Feng R, Chen X, Yu Y et al (2010) miR-126 functions as a tumour suppressor in human gastric cancer. Cancer Lett 298(1):50–63 Guo C, Sah JF, Beard L, Willson JK, Markowitz SD, Guda K (2008) The noncoding RNA, miR-126, suppresses the growth of neoplastic cells by targeting phosphatidylinositol 3-kinase signaling and is frequently lost in colon cancers. Genes Chromosomes Cancer 47(11):939–946 Tavazoie SF, Alarcón C, Oskarsson T et al (2008) Endogenous human microRNAs that suppress breast cancer metastasis. Nature 451(7175):147–152 Zhu N, Zhang D, Xie H et al (2011) Endothelial-specific intron-derived miR-126 is down-regulated in human breast cancer and targets both VEGFA and PIK3R2. Mol Cell Biochem 351(1–2):157–164 Huang X, Gschweng E, Van Handel B, Cheng D, Mikkola HK, Witte ON (2011) Regulated expression of microRNAs-126/126* inhibits erythropoiesis from human embryonic stem cells. Blood 117(7):2157–2165 Chodniewicz D, Klemke RL (2004) Regulation of integrin-mediated cellular responses through assembly of a CAS/Crk scaffold. Biochim Biophys Acta 1692(2–3):63–76 Gotoh T, Hattori S, Nakamura S et al (1995) Identification of Rap1 as a target for the Crk SH3 domain-binding guanine nucleotide-releasing factor C3G. Mol Cell Biol 15(12):6746–6753 Clark EA, Brugge JS (1995) Integrins and signal transduction pathways: the road taken. Science 268(5208):233–239 Birge RB, Knudsen BS, Besser D, Hanafusa H (1996) SH2 and SH3-containing adaptor proteins: redundant or independent mediators of intracellular signal transduction. Genes Cells 1(7):595–613 Becker CM, D’Amato RJ (2007) Angiogenesis and antiangiogenic therapy in endometriosis. Microvasc Res; 74:121–130 Xu YL, Wang DB, Liu QF, Chen YH, Yang Z (2010) Silencing of cofilin-1 gene attenuates biological behaviours of stromal cells derived from eutopic endometria of women with endometriosis. Hum Reprod 25(10):2480–2488 Crawford M, Brawner E, Batte K et al (2008) MiR-126 inhibits invasion in non-small cell lung carcinoma cell lines. Biochem Biophys Res Commun 373(4):607–612 Peltier HJ, Latham GJ (2008) Normalization of microRNA expression levels in quantitative RT-PCR assays: identification of suitable reference RNA targetsin normal and cancerous human solid tissues. RNA 14(5):844–852 Rhinn H, Marchand-Leroux C, Croci N, Plotkine M, Scherman D, Escriou V (2008) Housekeeping while brain’s storming Validation of normalizing factors for gene expression studies in a murine model of traumatic brain injury. BMC Mol Biol 9:62 Cicinnati VR, Shen Q, Sotiropoulos GC, Radtke A, Gerken G, Beckebaum S (2008) Validation of putative reference genes for gene expression studies in human hepatocellular carcinoma using real-time quantitative RT-PCR. BMC Cancer 8:350 Kitawaki J, Kado N, Ishihara H, Koshiba H, Kitaoka Y, Honjo H (2002) Endometriosis: the pathophysiology as an estrogen-dependent disease. J Steroid Biochem Mol Biol 83(1–5):149–155 Burney RO, Hamilton AE, Aghajanova L et al (2009) MicroRNA expression profiling of eutopic secretory endometrium in women with versus without endometriosis. Mol Hum Reprod 15(10):625–631 Pan Q, Luo X, Toloubeydokhti T, Chegini N (2007) The expression profile of micro-RNA in endometrium and endometriosis and the influence of ovarian steroids on their expression. Mol Hum Reprod 13(11):797–806 Schmidt M, De Mazière A, Smyczek T et al. (2007) The role of Egfl7 in vascular morphogenesis. Novartis Found Symp 283:18–28; discussion 28–36, 238–241 Schmidt M, Paes K, De Mazière A et al (2007) EGFL7 regulates the collective migration of endothelial cells by restricting their spatial distribution. Development 134(16):2913–2923 Harris TA, Yamakuchi M, Ferlito M, Mendell JT, Lowenstein CJ (2008) MiR-126 regulates endothelial expression of vascular cell adhesion molecule 1. Proc Natl Acad Sci USA 105(5):1516–1521 Mayer BJ, Hamaguchi M, Hanafusa H (1988) Characterization of p47gag-crk, a novel oncogene product with sequence similarity to a putative modulatory domain of protein-tyrosine kinases and phospholipase C. Cold Spring Harb Symp Quant Biol 53(Pt 2):907–914 Feller SM (2011) Crk family adaptors-signalling complex formation and biological roles. Oncogene 20(44):6348–6371 Watanabe T, Tsuda M, Tanaka S et al (2009) Adaptor protein Crk induces Src-dependent activation of p38 MAPK in regulation of synovial sarcoma cell proliferation. Mol Cancer Res 7(9):1582–1592 Ohlsson Teague EM, Van der Hoek KH, Hull LM et al (2009) MicroRNA-regulated pathways associated with endometriosis. Mol Endocrinol 23(2):265–275 Filigheddu N, Gregnanin I, Porporato PE et al (2010) Differential expression of microRNAs between eutopic and ectopic endometrium in ovarian endometriosis. J Biomed Biotechnol 2010:369549 Teague EM, Print CG, Hull ML (2010) The role of microRNAs in endometriosis and associated reproductive conditions. Hum Reprod Update 16(2):142–165 Noël JC, Chapron C, Borghese B, Fayt I, Anaf V (2011) Galectin-3 is Overexpressed in Various Forms of endometriosis. Appl Immunohistochem Mol Morphol 19(3):253–257 Vestergaard AL, Knudsen UB, Munk T, Rosbach H, Martensen PM (2011) Transcriptional expression of type-I interferon response genes and stability of housekeeping genes in the human endometrium and endometriosis. Mol Hum Reprod 17(4):243–254 Acknowledgments This research received support from the National Natural Science Foundation of China (No. 81070467), the Department of Science Technology of Liaoning Province, Shenyang, China (No.2009225026) and the Department of Science Technology of Shenyang, Shenyang, China (No.1091171-1-05). Conflict of interest We declare that we have no conflict of interest. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Liu, S., Gao, S., Wang, X.Y. et al. Expression of miR-126 and Crk in endometriosis: miR-126 may affect the progression of endometriosis by regulating Crk expression. Arch Gynecol Obstet 285, 1065–1072 (2012). https://doi.org/10.1007/s00404-011-2112-6 Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s00404-011-2112-6