NGF promotes mouse granulosa cell proliferation by inhibiting ESR2 mediated down-regulation of CDKN1A.
Nerve growth factor (NGF) is known to play key roles in ovarian follicular development, such as the assembly of early follicles and follicular ovulation through its high-affinity receptor, tyrosine kinase receptor A (trkA). Herein, the molecular mechanism controlling NGF-induced granulosa cell (GC) proliferation was not clear. In this study, we found that NGF is abundant in preantral GCs and knockdown of trkA in GCs attenuated NGF-induced GC proliferation and further decreased the levels of phosphorylated extracellular regulated protein kinases 1/2 (ERK1/2). Cyclin-dependent kinase inhibitor 1A (CDKN1A), also named p21, a factor which could be either a negative or a positive regulator via transformation related protein 53 (TRP53, also named p53)-dependent or independent pathways in cell proliferation, was up-regulated during the process of NGF-induced GC proliferation. Blockade of trkA (K252α) and ERK1/2 (U0126) in GCs decreased NGF-induced expression of CDKN1A and did not alter the expression of TRP53, indicating that NGF stimulates CDKN1A expression via the trkA-ERK1/2 pathway in a TRP53-independent manner. Meanwhile, ESR2, a tumor suppressor which is exclusively expressed in GCs, was suppressed in NGF-induced GC proliferation, and this effect was abrogated by U0126. Blockade of ESR2 (ICI182,780) caused the promotion of GC proliferation and CDKN1A expression, indicating that ESR2 may be downstream of the ERK1/2 pathway in mediating the effect of CDKN1A on NGF-induced GC proliferation. Therefore, ESR2 may be involved in the integration of intracellular signal cascades and cell cycle proteins in affecting GC proliferation. Here, we provide mechanistic insights into the roles of CDKN1A in NGF-induced GC proliferation. Understanding potential cross-points between CDKN1A and ESR2 affecting GC proliferation will help in the discovery of new therapeutic targets in some female infertility disorders.
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China.
Yong
Y
Wang
College of Life Science and Technology, Southwest University of Science and Technology, Mianyang, Sichuan 621010, China.
Wenjing
W
Liu
Xinxiang Medical College, Henan 453003, China.
Juan
J
Du
Jinzhou Medical College, Liaoning 121001, China.
Yang
Y
Yu
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China.
Ning
N
Liang
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China.
Meng
M
Liang
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China.
Guidong
G
Yao
China Agricultural University, Beijing 100094, China.
Sheng
S
Cui
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China; Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China.
Hefeng
H
Huang
International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China; Shanghai Key Laboratory of Reproductive Medicine, Shanghai 200025, China. Electronic address: sunfei@shsmu.edu.cn.
Fei
F
Sun
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects*; D047629:Estrogen Receptor beta / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D006107:Granulosa Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D008813:Mice, Inbred ICR; D009414:Nerve Growth Factors / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D020917:Receptor, trkA / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D050759:Cyclin-Dependent Kinase Inhibitor p21; D047629:Estrogen Receptor beta; D009414:Nerve Growth Factors; D016159:Tumor Suppressor Protein p53; D020917:Receptor, trkA; D048049:Extracellular Signal-Regulated MAP Kinases
CDKN1A (p21); Estrogen receptor beta (ESR2); Extracellular regulated protein kinases 1/2 (ERK1/2); Granulosa cells (GCs); Nerve growth factor (NGF)