APOM Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the APOM gene. This loss-of-function model avoids clonal selection bias, offering a heterogeneous population suitable for robust functional analyses of APOM-dependent processes.
The HT29 host cell line, originally derived from a primary colorectal adenocarcinoma, serves as a key model for human intestinal epithelial biology. Characterized by a near-diploid karyotype and adherent growth, these cells maintain the ability to differentiate upon appropriate stimuli, making them invaluable for studies of intestinal physiology, drug transport, and colorectal cancer. Their well-defined signaling networks provide an ideal platform for interrogating lipid-binding protein functions.
APOM, a lipocalin family member, functions as a chaperone for sphingosine-1-phosphate (S1P), stabilizing the lipid and presenting it to S1P receptors S1PR1, S1PR2, and S1PR3. Receptor engagement activates G-protein-coupled signaling cascades, notably PI3K/AKT and MAPK/ERK, which regulate eNOS, RhoA, and Rac1. APOM also associates with HDL apolipoproteins apoA-I and apoA-II and interacts with cubilin and megalin, participating in reverse cholesterol transport. Its expression is regulated by nuclear receptors including HNF-1??, HNF-4??, LXR, and FXR, as well as inflammatory cytokines TNF-?? and IL-1??.
Within HT29 intestinal epithelial cells, knockout of APOM disrupts the localized transport and signaling of S1P, leading to altered downstream effector activation including PI3K/AKT and MAPK/ERK cascades. This impairment influences critical cellular behaviors such as proliferation, migration, and barrier function, directly impinging on colorectal cancer aggressiveness and intestinal homeostasis. Furthermore, the loss of APOM-mediated interactions with HDL components permits dissection of lipid uptake and trafficking mechanisms relevant to systemic atherosclerotic processes and metabolic disorders.
Researchers can apply APOM Knockout HT29 Polyclonal Cells to study APOM/S1P signaling in colorectal cancer, employing assays like western blotting for APOM and phospho-AKT/ERK, RT-qPCR for S1PRs, and S1P quantification. HDL binding assays, co-immunoprecipitation with HDL components, and migration/proliferation studies support investigation of lipid transport and tumor cell behavior. The cells are also suitable for screening APOM-dependent modulators and modeling intestinal contributions to atherosclerosis, diabetes, sepsis, and liver disease. For further information, contact Ascent Research.
