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Articles by H. F Huang
Total Records ( 4 ) for H. F Huang
  Q Huang , A. P Cheung , Y Zhang , H. F Huang , N Auersperg and P. C. K. Leung

GDF-9 stimulates granulosa cell proliferation and plays important roles during folliclogenesis. However, its molecular mechanisms are still far from clear, particularly its roles in human granulosa cells around the periovulatory stage. Therefore, we investigated the effects of GDF-9 on cell cycle distribution, regulatory molecules, and signaling pathways involved in human luteinized granulosa (hLG) cells in vitro. Primary cultures of hLG cells obtained from women undergoing IVF and treated with and without recombinant GDF-9 were evaluated with and without a specific inhibitor to activin receptor-like kinase 5 (ALK5; SB-431542), ERK42/44 (PD-098059), or Smad3 (SIS3). Cell proliferation, cell cycle distribution, mRNA expression, and protein expression of relevant cell cycle molecules were determined by [3H]thymidine incorporation, flow cytometry, quantitative PCR, and immunoblotting, respectively. GDF-9 stimulated [3H]thymidine incorporation, enhanced cell transition from G0/G1 to S and G2/M phases (whereas both SB-431542 and PD-098059 attenuated these changes), increased mRNA and protein expression of cyclin D1 and E, and decreased those of the cyclin-dependent kinase (CDK) inhibitors p15INK4B and p16INK4A. GDF-9 also activated Rb protein (a critical G1 to S-phase regulator), ERK42/44, and Smad3. PD-098059 blocked Rb protein phorsphorylation and the increase in cyclin D1 and E but not the decrease in p15INK4B and p16INK4A induced by GDF-9. In contrast, SIS3 reversed the decrease in p15INK4B and p16INK4A but not the increase in cyclin D1 and E induced by GDF-9. GDF-9 stimulates hLG cell proliferation by stimulating cyclin D1 and E and suppressing p15INK4B and p16INK4A via both Smad-dependent and Smad-independent pathways.

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