AMF通过跨膜受体GPR30促进子宫内膜癌转移的机制研究

The traditional surgery plus chemotherapy/radiotherapy or existing targeted therapies have failed to significantly improve the survival rate of endometrial carcinoma which metastasis, suggesting other regulatory mechanisms may exist for endometrial cancer metastasis. Autocrine motility factor (AMF) mediated tumor cells migration、invasion and epithelial-mesenchymal transition (EMT); the rapid non-genomic effects induced by G protein-coupled receptor 30 (GPR30) promoted cancer cell invasion. Previous research have proved that：AMF and GPR30 can significantly increased tumor migration and invasion capacities in endometrial cancer，and AMF can up regulate the expression of membrane receptor GPR30. We presume and obtain preliminary validation of the preliminary experimental results: AMF can be bind with membrane receptor GPR30 in endometrial cancer cells, acting the MAPK-ERK signaling pathway to accelerate endometrial cancer invasion and distant metastasis. The present study aims to perform two gene modified mouse by CRISPR/Cas9 and TET-on system in female nude mouse for tumor-bearing, observe the effect of AMF on tumor biology in AMFR-null mice; analysis the AMF promoted endometrial cancer cells tumorigenicity through GPR30 in vivo; evaluate the feasibility of AMF as a prognostic marker in endometrial carcinoma. The research will deepen the understanding of the underlying mechanism of poorer prognosis of endometrial cancer, and provide a new approach of potential new molecular target for endometrial cancer progress and prognosis.

传统手术加放化疗、现有的靶向治疗未能显著改善转移内膜癌的生存率，提示内膜癌的转移存在其他调控机制。自分泌移动因子(AMF)可诱导肿瘤细胞迁移侵袭及上皮间充质转化（EMT）；GPR30可通过非基因组快速效应促进肿瘤细胞侵袭性。课题组前期研究证明: AMF及GPR30可显著增加内膜癌细胞迁移和侵袭的能力,此外AMF可调控GPR30表达。我们推测并得到预实验结果的初步验证:内膜癌中AMF与跨膜受体GPR30相互结合，激活下游MAPK-ERK信号通路促进肿瘤深层浸润和远处转移。课题将通过CRISPR/Cas9技术和TET-on诱导系统构建双基因修饰的原位移植瘤裸鼠模型,观察受体AMFR敲除后AMF对肿瘤生物学的影响;分析体内条件下AMF可通过GPR30增强肿瘤细胞的迁移侵袭能力。研究将会加深我们对转移性内膜癌凶险预后内在机制的理解,为寻找子宫内膜癌进展和预后的关键分子靶点提供新思路。

在国家自然科学基金委青年科学基金项目的资助下，本项目研究按计划执行，圆满完成了计划书中的研究内容：通过体内实验研究及临床组织标本验证,证实内膜癌中 AMF 可与跨膜受体 GPR30 相互结合，激活下游 MAPK-ERK 信号通路促进肿瘤细胞的迁移和侵袭，这种因子－受体相互结合产生的生物学效应促进内膜癌的发展。通过该研究，为内膜癌发生转移的机制提供重要理论及实验依据,进而为寻找能阻断肿瘤转移关键因子(GPR30, AMF)的分子靶向治疗提供可靠的转化研究基础。将有助于对远端转移及复发的内膜癌作出诊断。项目执行中，项目负责人学术水平不断得到提升，先后获得了上海市卫生计生系统优秀人才培养计划及国家自然科学基金委面上项目的资助。此外通过本项目的实施，联合培养了2名硕士研究生。