GNRH1 Knockout HCT 116 Polyclonal Cells consist of a CRISPR/Cas9?edited polyclonal population derived from the human colorectal carcinoma line HCT 116, engineered for targeted disruption of the GNRH1 gene. This polyclonal knockout pool contains diverse loss?of?function alleles, enabling functional analysis of gonadotropin?releasing hormone (GnRH) signaling without clonal selection bias. The cells provide a robust model for studying autocrine/paracrine GnRH effects in a cancer context.
The parental HCT 116 cell line carries an activating KRAS G13D mutation and is MLH1?deficient, resulting in mismatch repair deficiency and high genomic instability. This well?characterized colorectal carcinoma model exhibits active MAPK and PI3K signaling, making it suitable for investigating how oncogenic pathways intersect with neuropeptide?mediated regulation of proliferation, migration, and apoptosis.
GNRH1 encodes GnRH, a decapeptide that binds its receptor GNRHR, activating GNAQ/GNA11 and PLCB to mobilize calcium and activate PKC. This triggers MAPK1/3 (ERK1/2) phosphorylation, inducing transcription factors EGR1, FOS, and JUN to drive LHB and FSHB expression. Upstream, kisspeptin (KISS1) and neurokinin B (TAC3) stimulate GNRH1, while sex steroids provide feedback. In extrapituitary tissues, this pathway engages MAPK, cAMP, and calcium signaling to modulate cell behavior.
In HCT 116 cells, endogenous GnRH autocrine signaling may promote tumor progression. Knocking out GNRH1 disrupts this loop, allowing dissection of its role in KRAS?driven and mismatch repair?deficient colorectal cancer. The polyclonal format captures heterogeneous responses, enabling detection of subtle phenotypes in proliferation, invasion, and apoptosis assays, providing a clinically relevant tool to study crosstalk between reproductive hormone pathways and oncogenic signaling.
The polyclonal knockout cells are suitable for a variety of functional assays. Western blotting and RT?qPCR confirm GNRH1 disruption, while MTT and Transwell assays quantify changes in proliferation and migration. Phospho?ERK analysis and calcium imaging reveal signaling alterations. Reporter assays using LHB/FSHB promoters model hormonal regulation, and the model supports screening of GnRH analogs for anticancer activity. For more information, please contact Ascent Research.
