Presentation Authors: Zhenwei Han, Shijiazhuang, China, People's Republic of, Huiyang Xu*, Fu-Ju Chou, Yin Sun, Jiaqi Chen, Rochester, NY, Yong Zhang, Xiaolu Wang, Shijiazhuang, China, People's Republic of, Chawnshang Chang, Edward M. Messing, Shuyuan Yeh, Rochester, NY
Introduction: Recent studies suggested that ERÎ² could promote the ccRCC progression, and results from the TCGA database analyses indicate that ERÎ² expression is associated with the ccRCC stage, progression, and a worse overall survival. However, the detailed mechanisms by which ERÎ² regulates the RCC progression remains to be further elucidated. Our new finding indicates that ERÎ² could regulate circRNA to control ccRCC progression. Furthermore, early studies indicated that a subset of miRNAs were either up- or down-regulated in metastatic ccRCC, yet the functions of those RCC metastasis related miRNAs and how they interact with lncRNAs and circRNAs remain to be further elucidated.
Methods: RCC 786-O, A498, Caki-1 cells were used for the study. Lentiviral system was used for ectopic expression or knockdown of gene expressions. RNA extraction and quantitative Real-Time PCR were applied to determine the mRNA levels. Matrigel transwell and 3 D Invasion Assays were used to test the invasiveness of RCC cells. Chromatin Immunoprecipitation assay (ChIP) was applied to detect the protein- gene promoter (genomic DNA) binding. circRNA-miRNA pull-down assay was used to detect the interaction and function regulation between non-coding RNAs. Pre-clinical mouse RCC models were used to prove the therapeutic potential of FDA proved anti-estrogen or by targeting the ER-regulated signal pathways.
Results: Our data showed higher expressions or activities can promote ccRCC cell invasion. ERÎ² enhanced ccRCC cells invasion via altering the circATP2B1/miR-204-3p axis. This ERÎ²/circATP2B1/miR204-3p axis could promote ccRCC cell invasion via altering FN1 expression. Supportively, human clinical data show that higher ERÎ² expression is associated with the higher stage and poor prognosis in ccRCC patients. Importantly, preclinical study using in vivo mouse studies confirmed that ERÎ² promotes the ccRCC metastasis via altering the expression and function of circularRNA, circATP2B1.
Conclusions: Together, our study proved that ERÎ² promotes ccRCC cell invasion by altering the ERÎ²/circATP2B1/miR-204-3p/FN1 axis. These results further suggest that ERÎ² and circular RNA ATP2B1 may be applied as prognostic biomarkers for this disease, and that therapeutic targeting of this newly identified pathway may better prevent ccRCC progression.
Source of Funding: URMC Urology research fund and the George Whipple Professorship Endowment