The IL3 knockout UM-UC-3 polyclonal cells are a CRISPR/Cas9-edited population derived from the human bladder urothelial carcinoma cell line UM-UC-3, featuring targeted disruption of the IL3 gene. This polyclonal knockout product provides a heterogeneous pool of cells with diverse loss-of-function mutations, enabling comprehensive analysis of IL3-dependent signaling in a cancer context. CRISPR/Cas9-mediated gene inactivation preserves the native genetic background without selection markers, facilitating studies on pathways such as JAK-STAT, PI3K-AKT, and MAPK/ERK.
The parental UM-UC-3 cell line is a widely utilized model of invasive bladder urothelial carcinoma from a male patient, harboring mutant TP53 that drives its tumorigenic phenotype. UM-UC-3 cells are tumorigenic in immunocompromised mice, making them suitable for in vivo bladder cancer progression and therapeutic response studies. The IL3 knockout derivative maintains the core characteristics of the host line while specifically enabling investigation of IL3-mediated functions.
IL3 is a pleiotropic cytokine that primarily governs hematopoietic progenitor cell growth, but its signaling in non-hematopoietic tumors modulates the microenvironment. It activates JAK2 and STAT5 upon binding to IL3RA (CD123) and CSF2RB, and STAT5 transcriptionally upregulates BCL2, MYC, and CCND1 to promote survival and proliferation. IL3 also engages PI3K-AKT and MAPK/ERK pathways through GRB2, RAS, and RAF1. Upstream, IL3 expression is regulated by NFAT and AP-1 following TCR activation, IL-1, or TNF stimulation. IL3 knockout in these cells disrupts JAK2/STAT5 signaling, abolishing downstream anti-apoptotic and proliferative signals implicated in autocrine growth support and immune evasion in bladder tumors.
Loss of IL3 function in UM-UC-3 cells allows detailed examination of cytokine networks in urothelial carcinoma. The mutant p53 background facilitates cross-talk analysis between IL3 and p53 pathways. IL3 knockout may alter cytokine secretion affecting recruitment of IL3 receptor-expressing immune cells like mast cells and eosinophils. This model is highly relevant for studying tumor?Cstroma interactions, particularly in contexts of allergic inflammation and autoimmunity that mirror tumor immune dynamics.
Typical research applications include phospho-STAT5 flow cytometry and Western blotting for signaling analysis, RT-qPCR and ELISA for gene expression profiling, and cell proliferation assays to measure IL3-dependent growth. These polyclonal cells are suitable for drug sensitivity screening, co-culture chemotaxis assays, and in vivo xenograft studies to assess tumor growth. This versatile model supports mechanistic and translational bladder cancer research. For further information, contact Ascent Research.
