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IRS2 depletion inhibits cell proliferation and decreases hormone secretion in mouse granulosa cells

Lanjie Lei1, Feng Han1, Qiuyan Cui1

  • 1Clinical Skills Center, Affiliated Hospital of Jiujiang University, Jiujiang University, Jiangxi 332000, China.

The Journal of Reproduction and Development
|July 13, 2018
PubMed

Insights

Insulin receptor substrate 2 (IRS2) is crucial for mouse granulosa cell function. IRS2 knockdown impairs cell proliferation, steroidogenesis, and survival, highlighting its role in ovarian health via the PI3K/AKT pathway.

Area of Science:

  • Reproductive Biology
  • Cellular Signaling
  • Endocrinology

Background:

  • Insulin receptor substrate 2 (IRS2) is a key mediator in the insulin/insulin-like growth factor 1 (IGF1) signaling pathway.
  • IRS2 is known to be vital for hypothalamic and ovarian functions in mice.

Purpose of the Study:

  • To elucidate the specific roles of IRS2 in mouse granulosa cell (GC) steroidogenesis, apoptosis, cell cycle, and proliferation.
  • To investigate the underlying molecular mechanisms, including the involvement of the PI3K/AKT signaling pathway.

Main Methods:

  • IRS2 knockdown was performed in mouse GCs.
  • Cell proliferation and viability were assessed using CCK8 assays.
  • Apoptosis was analyzed via flow cytometry.
  • Gene and protein expression levels were measured (e.g., Cyclins, Bcl2, Bax, Star, Cyp11a1, Cyp19a1, Has2, Ptgs2).
  • Hormone concentrations (estradiol, progesterone) were quantified using ELISA.
  • PI3K/AKT pathway activation was evaluated.

Main Results:

  • IRS2 knockdown significantly inhibited GC proliferation, reduced cell viability, and increased apoptosis.
  • Key cell cycle regulators (Cyclin A1, B1) and anti-apoptotic protein (Bcl2) expression decreased, while pro-apoptotic (Bax) and other cell cycle proteins (Cyclin D1, D2) increased.
  • Steroidogenesis was impaired, evidenced by decreased estradiol (E2) and progesterone (P4) levels and reduced expression of key steroidogenic enzymes (Star, Cyp11a1, Cyp19a1).
  • Expression of folliculogenesis-related genes (Has2, Ptgs2) was altered.
  • IRS2's effects on cell viability and hormone secretion were dependent on the PI3K/AKT signaling pathway.

Conclusions:

  • IRS2 plays a critical role in regulating mouse granulosa cell proliferation and steroidogenesis.
  • The PI3K/AKT signaling pathway is essential for IRS2-mediated functions in GCs.
  • These findings provide insights into the molecular mechanisms governing ovarian function and potential targets for reproductive health research.
Keywords:
AKTCell proliferationGranulosa cellsInsulin receptor substrate 2 (IRS2)Steroidogenesis

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