HTRA1 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the UM-UC-3 human bladder transitional cell carcinoma cell line. This product provides a loss-of-function model for investigating the tumor suppressor role of the HTRA1 serine protease in urothelial carcinoma. The polyclonal nature ensures representation of diverse genetic edits, enabling robust phenotypic assessment without clonal isolation.
The UM-UC-3 host cell line was established from a male patient with bladder transitional cell carcinoma and exhibits epithelial adherent morphology. As a widely utilized in vitro model for urothelial carcinoma, UM-UC-3 cells retain key signaling pathways relevant to bladder cancer biology, including TGF-?? and Wnt signaling cascades. These cells serve as a relevant platform to interrogate mechanisms of tumor progression, migration, and invasion.
HTRA1 functions as a secreted serine protease that critically regulates TGF-?? signaling, apoptosis, and extracellular matrix (ECM) remodeling. Mechanistically, HTRA1 is transcriptionally activated by p53 and bone morphogenetic proteins (BMP2/4) in response to oxidative stress. It directly cleaves TGF-??1 and its receptors TGFBR1/TGFBR2, leading to reduced phosphorylation of SMAD2 and SMAD3 and diminished SMAD-dependent transcriptional activity. Concurrently, HTRA1 degrades ECM components such as fibronectin and modulates Wnt signaling by interacting with ??-catenin and TCF/LEF transcription factors, thereby influencing cell growth and adhesion.
In the context of bladder cancer, HTRA1 is frequently downregulated, and its loss is associated with enhanced TGF-?? signaling, increased cell migration, and invasive potential. The HTRA1 knockout in UM-UC-3 cells thus recapitulates a clinically relevant tumor suppressor disruption, permitting detailed dissection of HTRA1-dependent regulation of TGF-??1, SMAD2/3 phosphorylation, and downstream target gene expression. This model also enables exploration of crosstalk between TGF-?? and Wnt pathways mediated by ??-catenin, as well as HTRA1??s role in endoplasmic reticulum stress responses and insulin-like growth factor (IGF) signaling.
Researchers can utilize this polyclonal knockout population for a wide range of functional assays, including western blotting for HTRA1 and phospho-SMAD2, RT-qPCR analysis of TGF-?? target genes, cell migration and invasion assays, immunofluorescence to monitor SMAD localization, and cell viability assessments following TGF-?? pathway inhibition. Additional applications encompass RNA sequencing for global transcriptome profiling and ELISA to quantify secreted TGF-??1 levels. This product is also suitable for modeling HTRA1-related pathologies such as age-related macular degeneration and cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL), as well as for screening therapeutic compounds that target the TGF-?? axis. For comprehensive characterization data and experimental guidance, please contact Ascent Research.
