miR-4463 regulates aromatase expression and activity for 17β-estradiol synthesis in response to follicle-stimulating hormone

Objective The aim of this study was to investigate microRNAs (miRNAs) related to follicle-stimulating hormone (FSH) responsiveness using miRNA microarrays and to identify their target genes to determine the molecular regulatory pathways involved in FSH signaling in KGN cells. Methods To change the cellular responsiveness to FSH, KGN cells were treated with FSH receptor (FSHR)-specific small interfering RNA (siRNA) followed by FSH. miRNA expression profiles were determined through miRNA microarray analysis. Potential target genes of selected miRNAs were predicted using bioinformatics tools, and their regulatory function was confirmed in KGN cells. Results We found that six miRNAs (miR-1261, miR-130a-3p, miR-329-3p, miR-185-5p, miR-144-5p and miR-4463) were differentially expressed after FSHR siRNA treatment in KGN cells. Through a bioinformatics analysis, we showed that these miRNAs were predicted to regulate a large number of genes, which we narrowed down to cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and estrogen receptor alpha (ESR1) as the main targets for miR-4463. Functional analysis revealed that miR-4463 is a regulatory factor for aromatase expression and function in KGN cells. Conclusion In this study, we identified differentially expressed miRNAs related to FSH responsiveness. In particular, upregulation of miR-4463 expression by FSHR deficiency in human granulosa cells impaired 17β-estradiol synthesis by targeting CYP19A1 and ESR1. Therefore, our data might provide novel candidates for molecular biomarkers for use in research into poor responders.

[1]  W. Ni,et al.  FoxD2-AS1 promotes glioma progression by regulating miR-185-5P/HMGA2 axis and PI3K/AKT signaling pathway , 2019, Aging.

[2]  L. Yin,et al.  miR‐130a‐3p regulated TGF‐β1‐induced epithelial‐mesenchymal transition depends on SMAD4 in EC‐1 cells , 2019, Cancer medicine.

[3]  Bin Zhang,et al.  Long noncoding RNA FOXD2‐AS1 promotes glioma malignancy and tumorigenesis via targeting miR‐185‐5p/CCND2 axis , 2018, Journal of cellular biochemistry.

[4]  Xuemei He,et al.  Downregulation of MicroRNA-4463 Attenuates High-Glucose- and Hypoxia-Induced Endothelial Cell Injury by Targeting PNUTS , 2018, Cellular Physiology and Biochemistry.

[5]  Ghazal Haddad,et al.  miR-1266-5p and miR-185-5p Promote Cell Apoptosis in Human Prostate Cancer Cell Lines , 2018, Asian Pacific journal of cancer prevention : APJCP.

[6]  Y. Li,et al.  miR-106a Increases Granulosa Cell Viability and Is Downregulated in Women With Diminished Ovarian Reserve , 2018, The Journal of clinical endocrinology and metabolism.

[7]  Lin Fang,et al.  MiR-130a-3p inhibits migration and invasion by regulating RAB5B in human breast cancer stem cell-like cells. , 2018, Biochemical and biophysical research communications.

[8]  Xuemei He,et al.  MiR-4463 inhibits the migration of human aortic smooth muscle cells by AMOT , 2018, Bioscience reports.

[9]  Xuemei He,et al.  Roles of miR-4463 in H2O2-induced oxidative stress in human umbilical vein endothelial cells , 2017, Molecular medicine reports.

[10]  Jianhua Qiu,et al.  MiR-130a-3p inhibits the viability, proliferation, invasion, and cell cycle, and promotes apoptosis of nasopharyngeal carcinoma cells by suppressing BACH2 expression , 2017, Bioscience reports.

[11]  Wenfeng Li,et al.  MicroRNA-329-3p targets MAPK1 to suppress cell proliferation, migration and invasion in cervical cancer. , 2017, Oncology reports.

[12]  C. Leung,et al.  Circulating MicroRNAs in Delayed Cerebral Infarction After Aneurysmal Subarachnoid Hemorrhage , 2017, Journal of the American Heart Association.

[13]  A. Cupp,et al.  Gene expression profiling of bovine ovarian follicular and luteal cells provides insight into cellular identities and functions , 2017, Molecular and Cellular Endocrinology.

[14]  Y. Liu,et al.  Altered Plasma MicroRNAs as Novel Biomarkers for Arteriosclerosis Obliterans. , 2016, Journal of atherosclerosis and thrombosis.

[15]  Yanping Li,et al.  MicroRNA Expression is Altered in Granulosa Cells of Ovarian Hyperresponders , 2016, Reproductive Sciences.

[16]  A. Uyar,et al.  Poor ovarian response in women undergoing in vitro fertilization is associated with altered microRNA expression in cumulus cells. , 2015, Fertility and sterility.

[17]  I. Amiri,et al.  Estrogen and progesterone receptor subtype expression in granulosa cells from women with polycystic ovary syndrome , 2015, Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology.

[18]  Hui-min Hu,et al.  Circulating microRNAs in patients with polycystic ovary syndrome , 2015, Human fertility.

[19]  Yue Jiang,et al.  MicroRNA‐133b stimulates ovarian estradiol synthesis by targeting Foxl2 , 2013, FEBS letters.

[20]  K. Chaudhury,et al.  Oxidative status in granulosa cells of infertile women undergoing IVF , 2013, Systems biology in reproductive medicine.

[21]  Ying Zhou,et al.  Differentially expressed plasma microRNAs in premature ovarian failure patients and the potential regulatory function of mir-23a in granulosa cell apoptosis. , 2012, Reproduction.

[22]  A. Stavreus-Evers,et al.  Genetic predictors of controlled ovarian hyperstimulation: where do we stand today? , 2011, Human reproduction update.

[23]  Burton B. Yang,et al.  Micro-RNA378 (miR-378) regulates ovarian estradiol production by targeting aromatase. , 2011, Endocrinology.

[24]  L. Gianaroli,et al.  ESHRE consensus on the definition of 'poor response' to ovarian stimulation for in vitro fertilization: the Bologna criteria. , 2011, Human reproduction.

[25]  S. Kalantar,et al.  Investigating the association between polymorphism of follicle-stimulating hormone receptor gene and ovarian response in controlled ovarian hyperstimulation , 2011, Journal of human reproductive sciences.

[26]  Shalu Gupta,et al.  Demographic characteristics and clinical profile of poor responders in IVF / ICSI: A comparative study , 2010, Journal of human reproductive sciences.

[27]  J. Richards,et al.  The ovary: basic biology and clinical implications. , 2010, The Journal of clinical investigation.

[28]  Xin Li,et al.  MicroRNA-224 is involved in transforming growth factor-beta-mediated mouse granulosa cell proliferation and granulosa cell function by targeting Smad4. , 2010, Molecular endocrinology.

[29]  R. Behringer,et al.  The regulatory role of Dicer in folliculogenesis in mice , 2010, Molecular and Cellular Endocrinology.

[30]  M. Lalioti,et al.  Identification and in vitro characterization of follicle stimulating hormone (FSH) receptor variants associated with abnormal ovarian response to FSH. , 2010, The Journal of clinical endocrinology and metabolism.

[31]  D. Ovcharenko,et al.  Identification of MicroRNAs controlling human ovarian cell proliferation and apoptosis , 2009, Journal of cellular physiology.

[32]  N. Johnson,et al.  Ultrasound tests of ovarian reserve; A systematic review of accuracy in predicting fertility outcomes , 2009, Human fertility.

[33]  D. Seifer,et al.  Mullerian Inhibiting Substance is an ovarian growth factor of emerging clinical significance. , 2007, Fertility and sterility.

[34]  Xiaodong Cheng,et al.  Follicle-stimulating hormone induces multiple signaling cascades: evidence that activation of Rous sarcoma oncogene, RAS, and the epidermal growth factor receptor are critical for granulosa cell differentiation. , 2007, Molecular endocrinology.

[35]  He-feng Huang,et al.  Poor ovarian response to gonadotropin stimulation is associated with low expression of follicle-stimulating hormone receptor in granulosa cells. , 2007, Fertility and sterility.

[36]  B. Mol,et al.  A systematic review of tests predicting ovarian reserve and IVF outcome. , 2006, Human reproduction update.

[37]  S. Hammes,et al.  Ovarian Steroids: The Good, the Bad, and the Signals that Raise Them , 2006, Cell cycle.

[38]  R. Orvieto,et al.  Controlled ovarian hyperstimulation: Does prolonged stimulation justify cancellation of in vitro fertilization cycles? , 2005, Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology.

[39]  W. Rainey,et al.  Ovarian granulosa cell lines , 2004, Molecular and Cellular Endocrinology.

[40]  D. Maysinger,et al.  Perspectives on reproductive senescence and biological aging: studies in genetically altered follitropin receptor knockout [FORKO] mice , 2004, Experimental Gerontology.

[41]  Y. Nishi,et al.  Transrepression of Estrogen Receptor Signaling by Nuclear Factor-B in Ovarian Granulosa Cells , 2004 .

[42]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[43]  K. Britt,et al.  Estrogen actions in the ovary revisited. , 2002, The Journal of endocrinology.

[44]  E. Nieschlag,et al.  Isoforms and single nucleotide polymorphisms of the FSH receptor gene: implications for human reproduction. , 2002, Human reproduction update.

[45]  J. Gustafsson,et al.  Biological Role of Estrogen and Estrogen Receptors , 2002, Critical reviews in biochemistry and molecular biology.

[46]  E. Nieschlag,et al.  The Journal of Clinical Endocrinology & Metabolism Printed in U.S.A. Copyright © 2000 by The Endocrine Society Ovarian Response to Follicle-Stimulating Hormone (FSH) Stimulation Depends on the FSH Receptor Genotype* , 2022 .

[47]  G. Firestone,et al.  Follicle-Stimulating hormone (FSH) stimulates phosphorylation and activation of protein kinase B (PKB/Akt) and serum and glucocorticoid-lnduced kinase (Sgk): evidence for A kinase-independent signaling by FSH in granulosa cells. , 2000, Molecular endocrinology.

[48]  E. Nieschlag,et al.  The follicle-stimulating hormone receptor: biochemistry, molecular biology, physiology, and pathophysiology. , 1997, Endocrine reviews.

[49]  Z. Rosenwaks,et al.  Day 3 estradiol serum concentrations as prognosticators of ovarian stimulation response and pregnancy outcome in patients undergoing in vitro fertilization. , 1995, Fertility and sterility.

[50]  Y. Ito,et al.  Molecular basis of aromatase deficiency in an adult female with sexual infantilism and polycystic ovaries. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[51]  T. Bjøro,et al.  Ovarian stimulation in previous failures from in-vitro fertilization: distinction of two groups of poor responders. , 1990, Human reproduction.

[52]  K. Ryan.  Biochemistry of aromatase: significance to female reproductive physiology. , 1982, Cancer research.

[53]  R. Meuwissen,et al.  The role of microRNAs in biological processes. , 2014, Methods in molecular biology.

[54]  P. Devroey,et al.  Predictors of ovarian response: progress towards individualized treatment in ovulation induction and ovarian stimulation. , 2008, Human reproduction update.

[55]  F. Broekmans,et al.  Expected poor ovarian response in predicting cumulative pregnancy rates: a powerful tool. , 2008, Reproductive biomedicine online.

[56]  F. Slack,et al.  MicroRNAs in search of a target. , 2006, Cold Spring Harbor symposia on quantitative biology.

[57]  Y. Nishi,et al.  Transrepression of estrogen receptor beta signaling by nuclear factor-kappab in ovarian granulosa cells. , 2004, Molecular endocrinology.

[58]  Y. Nishi,et al.  Establishment and characterization of a steroidogenic human granulosa-like tumor cell line, KGN, that expresses functional follicle-stimulating hormone receptor. , 2001, Endocrinology.

[59]  Y. Nishi,et al.  Printed in U.S.A. Copyright © 2001 by The Endocrine Society Establishment and Characterization of a Steroidogenic Human Granulosa-Like Tumor Cell Line, KGN, That Expresses Functional Follicle-Stimulating Hormone Receptor , 2000 .