This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1703 human lung squamous cell carcinoma line, featuring targeted disruption of the ACER1 gene. ACER1 encodes alkaline ceramidase 1, which hydrolyzes ceramides to sphingosine and free fatty acids, playing a critical role in sphingolipid metabolism. The polyclonal nature reflects a heterogeneous population of edited cells, each carrying distinct CRISPR-induced modifications, making it suitable for studying gene function without clonal effects. This knockout model provides a stable loss-of-function system for investigating ceramide-mediated signaling in lung cancer research.
NCI-H1703 cells are a well-characterized human epithelial cell line derived from a lung squamous cell carcinoma. They represent a model of non-small cell lung cancer (NSCLC) with squamous histology, commonly used to study tumor biology, drug sensitivity, and signal transduction pathways. The cells retain key features of their malignant origin, including aberrant proliferation and resistance to apoptosis. Their genetic background and dependency on sphingolipid metabolism make them an ideal host for dissecting ACER1 function in the context of NSCLC.
ACER1 catalyzes the deacylation of ceramides, generating sphingosine and free fatty acids. Sphingosine is subsequently phosphorylated by sphingosine kinases (e.g., SPHK1) to sphingosine-1-phosphate (S1P), which signals through S1P receptors (S1PR1) to promote cell survival, proliferation, and migration. By controlling the balance between pro-apoptotic ceramides and pro-survival S1P, ACER1 acts as a critical rheostat. Knockout disrupts ceramide hydrolysis, leading to ceramide accumulation and reduced S1P production, thereby shifting the equilibrium toward apoptosis. Upstream regulators include TNF-alpha, IL-1beta, and cellular stress, which modulate ACER1 activity. Downstream, diminished S1P receptor signaling and altered crosstalk with ceramide synthases and sphingosine kinases reinforce the pro-apoptotic phenotype.
In NCI-H1703 cells, ACER1 disruption examines how ceramide accumulation and S1P deprivation influence oncogenic phenotypes. Elevated ceramides can potentiate apoptosis, potentially attenuating malignant characteristics. Conversely, decreased S1P impairs S1PR1-mediated survival pathways, including AKT and ERK activation. This model dissects endogenous ceramide metabolism in transformation and evaluates therapeutic targeting of sphingolipid enzymes. The interplay between ACER1 loss and inflammatory cytokines such as TNF-alpha and IL-1beta can be directly assessed, offering insights into lung cancer progression and treatment resistance.
Typical applications include mass spectrometry-based ceramide quantification and sphingolipidomics, apoptosis assays such as Annexin V/PI staining and western blotting for cleaved caspase-3, and cell proliferation studies. The model is ideal for investigating sphingolipid-mediated apoptosis regulation, drug resistance in NSCLC, and the impact of ceramide accumulation on oncogenic signaling cascades. It supports studies on how the ACER1-SPHK1-S1P axis integrates with TNF-alpha and IL-1beta signaling. For further inquiries regarding this product, please contact Ascent Research.
