METHODS: The potential interaction between E4 and GPER was investigated by molecular modeling and binding assays. The whole transcriptomic modulation triggered by E4 in TNBC cells via GPER was explored through high-throughput RNA sequencing analyses. Gene and protein expression evaluations as well as migration and invasion assays allowed us to explore the involvement of the GPER-mediated induction of the plasminogen activator inhibitor type 2 (SERPINB2) in the biological responses triggered by E4 in TNBC cells. Furthermore, bioinformatics analysis was aimed at recognizing the biological significance of SERPINB2 in ER-negative breast cancer patients.
RESULTS: After the molecular characterization of the E4 binding capacity to GPER, RNA-seq analysis revealed that the plasminogen activator inhibitor type 2 (SERPINB2) is one of the most up-regulated genes by E4 in a GPER-dependent manner. Worthy, we demonstrated that the GPER-mediated increase of SERPINB2 is engaged in the anti-migratory and anti-invasive effects elicited by E4 in TNBC cells. In accordance with these findings, a correlation between SERPINB2 levels and a good clinical outcome was found in ER-negative breast cancer patients.
CONCLUSIONS: Overall, our results provide new insights into the mechanisms through which E4 can halt migratory and invasive features of TNBC cells.
方法:通过分子建模和结合测定研究了E4与GPER之间的潜在相互作用。通过高通量RNA测序分析探索了经由GPER在TNBC细胞中由E4触发的整个转录组调节。基因和蛋白质表达评估以及迁移和侵袭测定使我们能够探索GPER介导的纤溶酶原激活物抑制剂2型(SERPINB2)在TNBC细胞中E4引发的生物反应中的参与。此外,生物信息学分析旨在认识SERPINB2在ER阴性乳腺癌患者中的生物学意义.
结果:在对E4与GPER的结合能力进行分子表征后,RNA-seq分析显示纤溶酶原激活物抑制剂2型(SERPINB2)是E4以GPER依赖性方式上调的基因之一。有价值的,我们证明GPER介导的SERPINB2增加参与了E4在TNBC细胞中引起的抗迁移和抗侵袭作用.根据这些发现,在ER阴性乳腺癌患者中发现SERPINB2水平与良好临床结局之间存在相关性.
结论:总体而言,我们的研究结果为E4阻止TNBC细胞迁移和侵袭性特征的机制提供了新的见解.