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原癌基因 c-erbB2 通过 PKC 和 MAPK 通路启动大鼠原始卵泡生长。

Proto-oncogene c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways.

机构信息

Department of Physiology Reproduction, Medical College of Nanchang University, Nanchang, China.

出版信息

Reprod Biol Endocrinol. 2010 Jun 19;8:66. doi: 10.1186/1477-7827-8-66.

DOI:10.1186/1477-7827-8-66
PMID:20565902
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2903600/
Abstract

BACKGROUND

c-erbB2, a proto-oncogene coding epidermal growth factor receptor-like receptor, also as a chemosensitivity/prognosis marker for gynecologic cancer, may be involved in initiation of growth of rat primordial follicles. The aim of the present study is to investigate the role and signal pathway of c-erbB2 in onset of rat primordial follicle development.

METHODS

The expression of c-erbB2 mRNA and protein in neonatal ovaries cultured 4 and 8 days with/without epidermal growth factor (EGF) were examined by in situ hybridization, RT-PCR and western blot. The function of c-erbB2 in the primordial folliculogenesis was abolished by small interfering RNA transfection. Furthermore, MAPK inhibitor PD98059 and PKC inhibitor calphostin were used to explore the possible signaling pathway of c-erbB2 in primordial folliculogenesis.

RESULTS

The results showed that c-erbB2 mRNA was expressed in ooplasm and the expression of c-erbB2 decreased after transfection with c-erbB2 siRNA. Treatment with EGF at 50 ng/ml significantly increased c-erbB2 expression and primary and secondary follicle formation in ovaries. However, this augmenting effect was remarkably inhibited by c-erbB2 siRNA transfection. Furthermore, folliculogenesis offset was blocked by calphostin (5 x 10(-4) mmol/L) and PD98059 (5 x 10(-2) mmol/L), but both did not down-regulate c-erbB2 expression. In contrast, the expressions of p-ERK and p-PKC were decreased obviously by c-erbB2 siRNA transfection.

CONCLUSIONS

c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways, suggesting an important role of c-erbB2 in rat primordial follicle initiation and development.

摘要

背景

c-erbB2 是一种原癌基因编码的表皮生长因子受体样受体，也是妇科癌症的化疗敏感性/预后标志物，可能参与大鼠原始卵泡生长的启动。本研究旨在探讨 c-erbB2 在大鼠原始卵泡发育中的作用及其信号通路。

方法

通过原位杂交、RT-PCR 和 Western blot 检测新生大鼠卵巢培养 4 天和 8 天，有/无表皮生长因子（EGF）时 c-erbB2mRNA 和蛋白的表达。用小干扰 RNA 转染法阻断 c-erbB2 的功能。此外，用 MAPK 抑制剂 PD98059 和 PKC 抑制剂 calphostin 探讨 c-erbB2 在原始卵泡发生中的可能信号通路。

结果

结果显示 c-erbB2mRNA 在卵质中表达，c-erbB2siRNA 转染后 c-erbB2 表达减少。50ng/mlEGF 处理可显著增加卵巢中 c-erbB2 的表达和初级和次级卵泡的形成。然而，这种增强作用被 c-erbB2siRNA 转染显著抑制。此外，calphostin（5×10（-4）mmol/L）和 PD98059（5×10（-2）mmol/L）阻断卵泡发生，但均不降低 c-erbB2 的表达。相反，c-erbB2siRNA 转染可明显降低 p-ERK 和 p-PKC 的表达。

结论

c-erbB2 通过 PKC 和 MAPK 途径启动大鼠原始卵泡生长，提示 c-erbB2 在大鼠原始卵泡启动和发育中起重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/776e805ae725/1477-7827-8-66-10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/304b1cb55ae1/1477-7827-8-66-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/c96dbfcbfd5f/1477-7827-8-66-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/876024a10340/1477-7827-8-66-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/0f42f10e30ab/1477-7827-8-66-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/6b676c64e44c/1477-7827-8-66-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/965b702d0eb1/1477-7827-8-66-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/860e95113f94/1477-7827-8-66-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/f2060b6559b7/1477-7827-8-66-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/990a28a723bd/1477-7827-8-66-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/776e805ae725/1477-7827-8-66-10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/304b1cb55ae1/1477-7827-8-66-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/c96dbfcbfd5f/1477-7827-8-66-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/876024a10340/1477-7827-8-66-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/0f42f10e30ab/1477-7827-8-66-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/6b676c64e44c/1477-7827-8-66-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/965b702d0eb1/1477-7827-8-66-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/860e95113f94/1477-7827-8-66-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/f2060b6559b7/1477-7827-8-66-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/990a28a723bd/1477-7827-8-66-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/148a/2903600/776e805ae725/1477-7827-8-66-10.jpg

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