Knockdown of the Premature Ovarian Insufficiency Candidate Gene NUP107 in Ovarian Granulosa Cells Affects Cell Functions, Including Receptor Expression and Estrogen Synthesis
REPRODUCTIVE AND DEVELOPMENTAL MEDICINE
Authors: Liu, Xiao-Cheng; Han, Meng-Xin; Xu, Yan; Wang, Hong-Yan; Li, Bin
Abstract
Objective: Mutations in NUP107 have been discovered in patients with premature ovarian insufficiency and may have tissue-specific effects in ovarian development. However, the role of NUP107 in human granulosa cell (GC) function and female fertility still remains unknown. In this study, we used RNA interference to investigate how NUP107 dysfunction influences GCs and ovarian development. Methods: Immunohistochemical staining was used to detect the expression of NUP107 in ovaries. Cell counting kit-8 assay, real-time cell analysis, and flow cytometry were used to explore cell proliferation and apoptosis, and an enzyme-linked immunosorbent assay was used to assess the estrogen concentrations. Quantitative real-time reverse transcription-quantitative polymerase chain reaction and Western blot analyses were used to determine the expression of NUP107 and functional receptors. Results: Knockdown of NUP107 expression had little effect on the growth and number of GCs. Further study confirmed that knockdown of NUP107 may interfere with estrogen synthesis in GCs and their sensitivity to the regulation of follicle-stimulating hormone (FSH) by decreasing the expression of estrogen synthesis-related genes AR, CYP17A1, CYP19A1, STAR, and NR5A1. Moreover, knockdown of NUP107 decreased the expression of AMHR2, FSHR, LHR, and ESR1 in GCs, but had no effect on the expression of ESR2. Conclusions: These data revealed that NUP107 may impede follicle growth and maturation by regulating hormone synthesis, sensitivity to follicle-stimulating hormone, and expression of functional receptors in GCs, and may, therefore, interfere with female fertility.
Concentration of activin A and follistatin in follicular fluid from human small antral follicles associated to gene expression of the corresponding granulosa cells
MOLECULAR AND CELLULAR ENDOCRINOLOGY
Authors: Jeppesen, J. V.; Nielsen, M. E.; Kristensen, S. G.; Andersen, C. Yding
Abstract
The present study correlated concentrations of activin A and follistatin in follicular fluid (FF) from human small antral follicles to FF concentrations of AMH, inhibin B, progesterone, and oestradiol and to the mRNA expression of FSH-receptor (FSHR), LH-receptor (LHR), AMH-receptor2 (AMHR2), CYP19a, and androgen-receptor (AR) in the corresponding granulosa cells (GC). FF from 144 follicles (3-12 mm in diameter) was included whereas mRNA expression profiles were established in GC from 66 of the 144 follicles. Levels of follistatin remained constant in relation to follicular diameter, whereas activin A levels increased in follicles exceeding 10 mm in diameter. Levels of activin A and inhibin B showed a highly significant inverse association. Follistatin showed highly significant positive associations with AMH and inhibin B levels and with FSHR and AR gene expression in GC. This study revealed unexpected associations that probably reflect the complicated regulatory mechanisms governing human folliculogenesis. (C) 2011 Elsevier Ireland Ltd. All rights reserved.
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