This CRISPR/Cas9-edited polyclonal GPER1 knockout cell population derives from the A2780 human ovarian carcinoma epithelial line, featuring targeted disruption of the GPER1 gene. The polyclonal format provides a heterogeneous pool of loss-of-function mutants, minimizing clonal artifacts and offering robust genetic ablation for functional studies. This model allows precise dissection of GPER1-dependent non-genomic estrogen signaling in ovarian cancer.
The A2780 cell line was established from an untreated endometrioid adenocarcinoma patient and is a standard ovarian cancer model. It expresses wild-type p53 and is cisplatin-sensitive, ideal for studying tumorigenesis and drug response. A2780 cells are widely used to examine estrogen-driven proliferation and migration in epithelial ovarian cancer.
Upon activation by 17??-estradiol, GPER1 couples to G??s, G??i/o, and G??q, initiating parallel signaling cascades. The receptor triggers Src-mediated MMP activation, releasing HB-EGF to transactivate EGFR, which subsequently recruits adaptor proteins and activates the Ras-Raf-MEK-ERK kinase cascade and PI3K/AKT signaling. Simultaneously, G??q drives PLC-IP3-mediated calcium flux, while G??s stimulates cAMP production activating PKA and the transcription factor CREB. These pathways converge to upregulate immediate-early genes such as c-fos, cell cycle drivers like cyclin D1, and anti-apoptotic Bcl-2. GPER1 signaling is modulated by interacting proteins including ??-arrestin, which scaffolds signalosomes, and caveolin, which organizes membrane microdomains. The receptor also responds to estrone, aldosterone, xenoestrogens, and the partial agonist tamoxifen, integrating hormonal and growth factor inputs.
In A2780 ovarian cancer cells, GPER1 promotes estrogen-induced proliferation and migration. CRISPR-mediated knockout eliminates receptor function, allowing clean dissection of GPER1-specific roles independent of nuclear ERs. Because A2780 cells are cisplatin-sensitive and express wild-type p53, the knockout model is instrumental for studying how GPER1 influences chemoresistance. Tamoxifen’s partial agonism at GPER1 can be examined in this background to clarify its off-target effects. Furthermore, loss of GPER1 may reveal compensatory signaling, providing insights into resistance to endocrine therapies.
These polyclonal knockout cells are amenable to diverse assays. Western blot detection of phospho-ERK and phospho-Akt, and RT-qPCR for c-fos and cyclin D1, quantify pathway activation. Proliferation (MTT), migration/invasion (Transwell), and calcium mobilization assays assess functional responses. Co-immunoprecipitation examines GPER1-EGFR complexes, while flow cytometry enables cell cycle profiling. Cisplatin sensitivity tests evaluate chemoresistance. The polyclonal population also facilitates ligand screening for selective GPER1 modulators. For more information, contact Ascent Research.
