The HTRA1 Knockout KYSE-150 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal human cell population with targeted disruption of the HTRA1 gene. This polyclonal knockout product provides a heterogeneous loss-of-function model, avoiding single-cell cloning bias while preserving the inherent genetic diversity of the host cell background. These cells enable the study of HTRA1-dependent processes within an esophageal squamous carcinoma context, using CRISPR/Cas9-mediated gene disruption.
The parental KYSE-150 cell line is an epithelial model derived from a poorly differentiated human esophageal squamous cell carcinoma. This well-characterized line retains critical signaling pathways, including TGF-?? and MAPK/ERK cascades, and is widely used to investigate the molecular mechanisms of esophageal cancer pathogenesis, metastasis, and therapy resistance.
HTRA1 is a secreted serine protease that suppresses tumorigenesis by cleaving TGF-?? precursors and extracellular matrix proteins such as decorin (DCN) and fibronectin (FN1). This proteolytic activity inhibits TGF-?? signaling, leading to reduced phosphorylation of AKT1, MAPK3/ERK1, and MAPK1/ERK2, and promotes caspase-3 (CASP3)-mediated apoptosis. HTRA1 expression is controlled by upstream regulators including TGFB1, EGF, FGF2, TP53, HIF1A, and epigenetic methylation. Its network intersections with SERPINA1 further modulate proteolytic balance. Collectively, HTRA1 acts as a pivotal negative regulator of the TGF-??/SMAD, PI3K/AKT, and MAPK/ERK pathways.
In esophageal squamous cell carcinoma, HTRA1 is frequently epigenetically silenced, contributing to enhanced TGF-?? activity, sustained AKT and ERK signaling, and apoptotic resistance. This polyclonal HTRA1 knockout model in KYSE-150 cells mirrors this loss-of-function scenario, facilitating investigation of how HTRA1 deficiency drives proliferation, migration, and invasion. The model is instrumental for exploring the crosstalk between extracellular matrix remodeling and oncogenic kinase signaling in esophageal carcinoma biology.
These cells are applicable in diverse functional assays. They support western blotting and RT-qPCR for analyzing HTRA1 downstream targets, Transwell migration and invasion assays, Annexin V apoptosis measurement, and TGF-?? pathway reporter analyses. Further applications include co-immunoprecipitation of HTRA1 interactors, immunofluorescence localization, and cell proliferation studies. The model also serves drug screening for HTRA1 reactivation strategies. For additional information, contact Ascent Research.
