The G Protein Estrogen Receptor

• Published in: Journal of experimental & clinical cancer research 2024-06, Vol.43 (1)
• Main Authors: Zicarelli, Azzurra, Occhiuzzi, Maria Antonietta, Rizzuti, Bruno, Talia, Marianna, De Rosis, Salvatore, De Francesco, Ernestina Marianna, Maggiolini, Marcello, Belfiore, Antonino, Greco, Gianluigi, Grande, Fedora, Guzzo, Antonella, Fortino, Giancarlo, Cirillo, Francesca, Lappano, Rosamaria, Scordamaglia, Domenica, Miglietta, Anna Maria, Capalbo, Carlo, Di Dio, Marika
• Format: Article
• Language: English
• Subjects: Analysis; Antimitotic agents; Antineoplastic agents; Breast cancer; Care and treatment; Diagnosis; Dosage and administration; Epidermal growth factor; G proteins
• ISSN: 0392-9078

Background The cyclin D1-cyclin dependent kinases (CDK)4/6 inhibitor palbociclib in combination with endocrine therapy shows remarkable efficacy in the management of estrogen receptor (ER)-positive and HER2-negative advanced breast cancer (BC). Nevertheless, resistance to palbociclib frequently arises, highlighting the need to identify new targets toward more comprehensive therapeutic strategies in BC patients. Methods BC cell lines resistant to palbociclib were generated and used as a model system. Gene silencing techniques and overexpression experiments, real-time PCR, immunoblotting and chromatin immunoprecipitation studies as well as cell viability, colony and 3D spheroid formation assays served to evaluate the involvement of the G protein-coupled estrogen receptor (GPER) in the resistance to palbociclib in BC cells. Molecular docking simulations were also performed to investigate the potential interaction of palbociclib with GPER. Furthermore, BC cells co-cultured with cancer-associated fibroblasts (CAFs) isolated from mammary carcinoma, were used to investigate whether GPER signaling may contribute to functional cell interactions within the tumor microenvironment toward palbociclib resistance. Finally, by bioinformatics analyses and k-means clustering on clinical and expression data of large cohorts of BC patients, the clinical significance of novel mediators of palbociclib resistance was explored. Results Dissecting the molecular events that characterize ER-positive BC cells resistant to palbociclib, the down-regulation of ER[alpha] along with the up-regulation of GPER were found. To evaluate the molecular events involved in the up-regulation of GPER, we determined that the epidermal growth factor receptor (EGFR) interacts with the promoter region of GPER and stimulates its expression toward BC cells resistance to palbociclib treatment. Adding further cues to these data, we ascertained that palbociclib does induce pro-inflammatory transcriptional events via GPER signaling in CAFs. Of note, by performing co-culture assays we demonstrated that GPER contributes to the reduced sensitivity to palbociclib also facilitating the functional interaction between BC cells and main components of the tumor microenvironment named CAFs. Conclusions Overall, our results provide novel insights on the molecular events through which GPER may contribute to palbociclib resistance in BC cells. Additional investigations are warranted in order to assess whether targeting th