The EHHADH Knockout HGC-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population in which the EHHADH gene has been disrupted in the HGC-27 human gastric carcinoma cell line. EHHADH encodes a bifunctional peroxisomal enzyme possessing both enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activities, essential for the beta-oxidation of long-chain fatty acids within peroxisomes. This polyclonal knockout pool comprises a heterogeneous mixture of cells harboring various CRISPR-induced loss-of-function alleles, providing a versatile and robust model for studying the role of peroxisomal fatty acid oxidation in cancer biology and metabolic disease. The use of a polyclonal population mitigates clonal variation and offers a realistic representation of gene disruption effects in a cellular context.
The host cell line HGC-27 is an undifferentiated gastric adenocarcinoma epithelial cell line originally derived from the lymph node metastasis of a patient with gastric carcinoma. These cells are widely employed as a model system for metastatic gastric cancer, exhibiting aggressive growth characteristics and genetic alterations typical of advanced gastric malignancies. HGC-27 cells are particularly valuable for investigating the molecular mechanisms underlying tumor invasion, metastasis, and metabolic adaptations, including alterations in lipid metabolism that support cancer cell survival and proliferation.
EHHADH functions within the peroxisomal fatty acid beta-oxidation pathway, catalyzing the second and third steps of the spiral: hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA and subsequent oxidation to 3-ketoacyl-CoA, with concomitant reduction of NAD+ to NADH. The enzyme operates in concert with other pathway components such as the peroxisomal ABC transporter ABCD1, acyl-CoA oxidase ACOX1, acetyl-CoA acyltransferase ACAA1, and sterol carrier protein SCP2. Its expression is transcriptionally regulated by the nuclear receptor PPARA and its coactivator PPARGC1A, which are activated by fatty acid ligands. Peroxisomal import of EHHADH depends on the receptor PEX5, linking its localization to peroxisomal biogenesis. Downstream, the reaction products??acetyl-CoA, medium-chain acyl-CoAs, and NADH??feed into energy production and biosynthetic pathways, highlighting the metabolic significance of EHHADH.
In the context of gastric cancer, metabolic reprogramming is a hallmark, with cancer cells often relying on fatty acid oxidation to meet energetic and biosynthetic demands, particularly under nutrient-deprived conditions or during metastasis. The HGC-27 cell line, being metastatic in origin, provides a clinically relevant background to explore how peroxisomal fatty acid oxidation contributes to gastric cancer progression. Disruption of EHHADH is expected to impair peroxisomal beta-oxidation, leading to accumulation of very long-chain fatty acids and reduced generation of energy-rich metabolites, which may sensitize cells to metabolic stress and impact tumorigenic properties.
This EHHADH knockout polyclonal cell product is rigorously validated for gene disruption and is suitable for a range of functional analyses, including western blotting to assess protein expression, RT-qPCR to confirm transcript reduction, fatty acid oxidation assays to measure metabolic flux, peroxisomal staining to evaluate organelle integrity, lipidomics profiling to detect fatty acid accumulation, and cellular respiration assays to monitor metabolic consequences. It serves as a powerful tool for investigating peroxisomal fatty acid oxidation in gastric cancer, identifying metabolic vulnerabilities for therapeutic intervention, and modeling peroxisomal disorders. For additional details or to inquire about customized gene-editing services, please contact Ascent Research.
