Autocrine motility factor promotes endometrial cancer progression by targeting GPER-1
Autocrine motility factor promotes endometrial cancer progression by targeting GPER-1
复制标题
自分泌运动因子通过靶向 GPER-1 促进子宫内膜癌进展
DOI：
10.1186/s12964-019-0336-4
复制
发表时间：
2019-03-05
影响因子：
8.4
通讯作者：
Wan, Xiao-Ping
中科院分区：
文献类型：
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作者：
Li, Yiran;Jia, Yuanhui;Wan, Xiao-Ping
背景自分泌运动因子（AMF）是调节子宫内膜癌（EC）侵袭性的重要因子。多项证据表明，某些生长因子通过G蛋白偶联雌激素受体（GPER）信号通路促进肿瘤细胞的迁移和增殖。本研究的目的是探讨GPER-1在AMF介导的EC复发和进展的调节机制中的作用。方法采用实时细胞分析（RTCA）方法，研究AMF是否依赖于EC细胞中的自分泌运动因子受体（AMFR）信号。利用全基因组表达谱芯片和酵母双杂交技术检测AMF和GPER-1在AMFR缺失情况下的相互作用，并通过免疫共沉淀和免疫荧光实验证实两者的物理相互作用。使用相对和绝对定量的同位素标签（iTRAQ）分析来鉴定由AMF-GPER-1相互作用激活的靶途径。用或不用外源性AMF处理携带源自经修饰的SPEC 2细胞系的异种移植物的小鼠群，以验证先前实验的结果。免疫组化进行评估AMF和GPER-1的表达在子宫内膜癌标本和正常endometrium.ResultsOur数据表明，GPER-1结合AMF和形成的复合物从质膜易位到细胞质。机制研究表明，AMF与GPER-1之间的相互作用触发磷酸肌醇-3-激酶信号传导并促进EC细胞生长。更重要的是，通过动物实验和人体组织实验，我们发现AMF参与了GPER-1介导的EC进展，这与上述观察结果相一致。结论我们的工作不仅阐明了AMF-GPER-1-AKT信号级联对子宫内膜癌进展的调控机制，而且为靶向该通路治疗子宫内膜癌奠定了基础。
BackgroundAutocrine motility factor (AMF) is a critical factor regulating aggressiveness of endometrial cancer (EC). Multiple pieces of evidence indicate that it is through G protein coupled estrogen receptor (GPER) signaling pathway that some growth factors promoted the migration and proliferation of tumor cells. The aim of this study is to explore the role of GPER-1 in AMF mediated regulatory mechanisms of EC recurrence and progression.MethodsReal-Time Cell Analysis (RTCA) assays were performed to assess whether AMF depends on Autocrine motility factor recepter (AMFR) signaling in EC cells. A genome-wide expression microarray and Yeast Two-Hybrid assay were used to detect AMF and GPER-1 interaction in the context of AMFR depletion, and co-immunoprecipitation and immunofluorescence experiments were performed to confirm the physical interaction. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) analysis was used for the identification of the target pathway activated by AMF-GPER-1 interaction. Cohorts of mice harboring xenografts derived from modified SPEC2 cell lines were treated with or without exogenous AMF to validate the results of previous experiments. Immunohistochemistry was performed to assess AMF and GPER-1 expression in endometrial cancer specimens and normal endometrium.ResultsOur data showed that GPER-1 binds to AMF and the formed complex translocates from the plasma membrane to the cytoplasm. Mechanistic investigations demonstrated that interaction between AMF and GPER-1 triggers phosphoinositide-3-kinase signaling and promotes EC cell growth. More importantly, through animal experiments and human tissue experiments, we found that AMF contributes to GPER-1-mediated EC progression, which is consistent with the above observations.ConclusionsOur work not only delineated the regulatory mechanisms of endometrial cancer progression by AMF-GPER-1-AKT signaling cascade but also laid the foundation of targeting this pathway for treating endometrial cancer.