The HRH1 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human ovarian adenocarcinoma cell line A2780, with targeted disruption of the HRH1 gene. This loss-of-function model abolishes histamine H1 receptor signaling, providing a tool to dissect histamine-dependent pathways in a cancer-relevant context. The polyclonal population preserves the heterogeneous genetic background of the edited pool, avoiding clonal selection and enabling studies that require population-level responses, such as functional genomics and drug target validation.
The A2780 cell line is a widely used human ovarian carcinoma epithelial cell line originally established from an untreated patient with ovarian adenocarcinoma. It serves as a standard model for ovarian cancer research, reflecting key characteristics of epithelial ovarian tumors, and is commonly employed in investigations of cell proliferation, apoptosis, drug resistance, and hormone signaling. This host cell line provides a clinically pertinent platform for examining histamine-mediated mechanisms in ovarian cancer biology.
HRH1 encodes the histamine H1 receptor, a Gq/11-coupled G-protein-coupled receptor. Upon histamine binding, it activates phospholipase C-beta (PLC-beta), which hydrolyzes PIP2 into inositol trisphosphate (IP3) and diacylglycerol (DAG). IP3 mobilizes intracellular calcium, while DAG activates protein kinase C (PKC), leading to downstream mitogen-activated protein kinase (MAPK) signaling and nuclear factor-kB (NF-kB) activation. HRH1 knockout eliminates this signaling cascade, preventing calcium flux, PKC activation, and NF-kB-mediated transcription of pro-inflammatory targets such as IL8 and PTGS2. The receptor is upregulated by TNF-alpha and IL-1beta and desensitized by beta-arrestin and G-protein-coupled receptor kinases (GRKs).
In A2780 ovarian cancer cells, histamine/HRH1 signaling modulates proliferation, migration, and secretion of inflammatory cytokines. Disruption of HRH1 allows interrogation of its specific contributions to tumor cell behavior and the inflammatory microenvironment. The knockout abrogates the Gq/11?CPLC-beta?CCa2+?CPKC?CNF-kB axis, reducing expression of NF-kB-dependent genes and potentially impacting angiogenesis and immune cell recruitment. This model is valuable for distinguishing cancer cell-autonomous effects from paracrine signals in co-culture systems and for studying HRH1 involvement in chemoresistance.
These polyclonal knockout cells support diverse experimental approaches including Western blotting for protein expression analysis, calcium flux assays to measure histamine-induced mobilization, NF-kB reporter assays, and ELISA for IL-8. Proliferation, migration, and invasion assays provide functional readouts, while RT-qPCR quantifies target gene expression. Flow cytometry verifies receptor loss. The model is ideal for GPCR pharmacology, allergy and inflammation research, and drug screening in ovarian cancer. For technical inquiries, contact Ascent Research.
