nm23 regulates decidualization through the PI3K-Akt-mTOR signaling pathways in mice and humans
nm23 regulates decidualization through the PI3K-Akt-mTOR signaling pathways in mice and humans
复制标题
nm23 通过 PI3K-Akt-mTOR 信号通路调节小鼠和人类的蜕膜化
DOI：
10.1093/humrep/dew191
复制
发表时间：
2016-10-01
影响因子：
6.1
通讯作者：
Ding，Yu-Bin
中科院分区：
文献类型：
--
作者：
Zhang，Xue;Fu，Li-Juan;Ding，Yu-Bin
研究问题nm 23在小鼠和人类的蜕膜化中是否具有功能意义？综述nm 23通过磷酸肌醇3激酶/哺乳动物雷帕霉素靶蛋白（PI 3 K-Akt-mTOR）信号通路影响小鼠子宫内膜基质细胞（ESCs; mESCs）和人ESCs的蜕膜化。已知nm 23在抑制转移中的功能已在多种癌症类型中得到证实。nm 23还参与DNA复制和细胞增殖与分化的控制。研究设计、大小和持续时间我们首先分析了nm 23在小鼠妊娠早期（n= 6/组）、假孕（n= 6/组）和人工蜕膜化（n = 6/组）以及人类月经周期阶段和妊娠早期的表达谱。然后，我们使用原代培养的胚胎干细胞和人胚胎干细胞系，T-HESC，探讨nm 23的激素调节和nm 23 invitrodecidualization的作用，并作为下游PI 3 K-Akt-mTOR信号转导pathways.PARTICIPANTS/MATERIALS，SETTINGS AND METHODS我们评估了nm 23在小鼠和人类的动态表达，免疫组化，蛋白质印迹和实时定量RT-PCR（RT-qPCR）。通过蛋白质印迹在分离的原代mESC和T-HESC中研究类固醇激素对nm 23的调节。通过5-乙炔基-2 ′-脱氧尿苷染色（EdU）和增殖细胞核抗原蛋白（PCNA）表达分析nm 23基因敲低（siRNA）对ESC增殖的影响。采用小鼠分化标志物蜕膜/滋养层PRL相关蛋白（dtprp，也称为prl 8a 2）和催乳素家族3亚家族c成员1（prl 3c 1）以及人分化标志物胰岛素样生长因子结合蛋白1（IGFBP 1）和催乳素（PRL），通过RT-qPCR测定nm 23表达对ESCs分化的影响。Western blot检测nm 23 siRNA（si-nm 23）和PI 3 K抑制剂LY 294002对nm 23下游PI 3 K-Akt-mTOR信号通路的影响。主要结果及作用CHANCENM 23-M1在早孕蜕膜区和人工诱导的蜕膜瘤中特异性表达，NM 23-H1在人早孕蜕膜中强表达。nm 23的表达在体外mESC和T-HESC中被雌二醇和孕酮上调（与对照相比P< 0.05），并且这被它们各自的受体拮抗剂ICI 182、780和RU 486抑制。小鼠和人nm 23基因敲低降低ESC的增殖和分化（P< 0.05）。nm 23是否通过激活AMPK、RAS、PKA、STAT 3或其他信号分子来调控蜕膜化，目前尚不清楚。nm 23在蜕膜化中的作用是在体外测试的。结果表明，nm 23在小鼠和人类的蜕膜化中起着至关重要的作用，nm 23基因的表达是受卵巢调节的。nm 23在早孕期蜕膜中的下调可能与女性不孕症有关。研究基金/竞争兴趣（S）本研究由国家自然科学基金资助。
STUDY QUESTIONDoes nm23 have functional significance in decidualization in mice and humans?SUMMARY ANSWERnm23 affects decidualization via the phosphoinositide 3 kinase/mammalian target of rapamycin (PI3K-Akt-mTOR) signaling pathways in mouse endometrial stromal cells (ESCs; mESCs) and human ESCs.WHAT IS KNOWN ALREADYThe function of nm23 in suppressing metastasis has been demonstrated in a variety of cancer types. nm23 also participates in the control of DNA replication and cell proliferation and differentiation.STUDY DESIGN, SIZE AND DURATIONWe first analyzed the expression profile of nm23 in mice during early pregnancy (n= 6/group), pseudopregnancy (n= 6/group) and artificial decidualization (n= 6/group) and in humans during the menstrual cycle phases and the first trimester. We then used primary cultured mESCs and a human ESC line, T-HESC, to explore the hormonal regulation of nm23 and the roles of nm23 inin vitrodecidualization, and as a possible mediator of downstream PI3K-Akt-mTOR signaling pathways.PARTICIPANTS/MATERIALS, SETTINGS AND METHODSWe evaluated the dynamic expression of nm23 in mice and humans using immunohistochemistry, western blot and real-time quantitative RT-PCR (RT-qPCR). Regulation of nm23 by steroid hormones was investigated in isolated primary mESCs and T-HESCs by western blot. The effect of nm23 knockdown (using siRNA) on ESC proliferation was analyzed by 5-ethynyl-2′-deoxyuridine staining (EdU) and proliferating cell nuclear antigen protein (PCNA) expression. The influence of nm23 expression on the differentiation of ESCs was determined by RT-qPCR using the mouse differentiation markers decidual/trophoblast PRL-related protein (dtprp, also named prl8a2) and prolactin family 3 subfamily c member 1 (prl3c1) and the human differentiation markersinsulin-like growth factor binding protein 1(IGFBP1) andprolactin(PRL). The effects of nm23 siRNA (si-nm23) and the PI3K inhibitor LY294002 on the downstream effects of nm23 on the PI3K-Akt-mTOR signaling pathway were estimated by western blot.MAIN RESULTS AND THE ROLE OF CHANCENM23-M1 was specifically expressed in the decidual zone during early pregnancy and in artificially induced deciduoma, and NM23-H1 was strongly expressed in human first trimester decidua. The expression of nm23 was upregulated by oestradiol and progesterone (P< 0.05 versus control)in vitroin mESCs and T-HESC, and this was inhibited by their respective receptor antagonists, ICI 182,780 and RU486. Mouse and human nm23 knockdown decreased ESC proliferation and differentiation (P< 0.05 versus control). The PI3K-Akt-mTOR signaling pathways were downstream mediators of nm23 in mESCs and T-HESCs decidualization.LIMITATIONS AND REASONS FOR CAUTIONWhether the nm23 regulates decidualization via the activation of AMPK, RAS, PKA, STAT3 or other signaling molecules remains to be determined. The role of nm23 in decidualization was testedin vitroonly.WIDER IMPLICATIONS OF THE FINDINGSResults demonstrate that nm23 plays a vital role in decidualization in mice and humans and that nm23 gene expression is hormonally regulated. The downregulation of nm23 in decidua during the first trimester may be associated with infertility in women.STUDY FUNDING/COMPETING INTEREST(S)This study was supported by the National Natural Science …