Objective: Premature ovarian failure, a reproductive dysfunction characterized by follicle loss leads to premature menopause. Apoptosis of granulosa cells may be responsible for the associated follicle depletion. MicroRNAs are expressed abundantly in granulosa cells and play an important role in follicular atresia. Evidence suggests that signal transducer and activator of transcription 3 (STAT3) is involved in follicle growth and female fertility.
Methods: We incubated cultured mouse granulosa cells (mGCs) with increasing doses of cisplatin (CP) for varying periods. Cell proliferation and apoptosis were measured by Cell Counting Kit-8 assay, flow cytometry, and protein expression of cleaved caspase-3. Western blot analysis was used to assess STAT3 and phospho-STAT3 after mGCs were transfected with a microRNA-125a-5p (miR-125a-5p) mimic and a miR-125a-5p inhibitor, respectively. Luciferase reporter assay was conducted to determine the relationship between miR-125a-5p and STAT3.
Results: CP reduced mGC viability, progesterone levels, and estradiol levels. miR-125a-5p was up-regulated in CP-treated mGCs, whereas STAT3 was down-regulated. Increased apoptosis and cleaved caspase-3 were observed in mGCs transfected with a miR-125a-5p mimic or STAT3 interference fragment. Protein expression of STAT3 and phospho-STAT3 was up-regulated or down-regulated when transfected with a miR-125a-5p inhibitor or miR-125a-5p mimic, respectively. Luciferase reporter assays indicated that miR-125a-5p targets the 3' untranslated region of STAT3.
Conclusions: Overexpression of miR-125a-5p promotes mGC apoptosis by targeting STAT3. Our findings imply the important role of miR-125a-5p in the pathogenesis of premature ovarian failure.