The EHHADH Knockout MES-OV Polyclonal Cells constitute a human ovarian cancer model engineered with CRISPR/Cas9-mediated disruption of the EHHADH gene, producing a heterogeneous polyclonal knockout cell population. This polyclonal product provides a stable loss-of-function system for investigating peroxisomal fatty acid ??-oxidation and its roles in cancer cell metabolism. The gene editing generates a pool of cells with varied EHHADH disruption, enabling robust functional studies without reliance on a single clonal isolate.
The host cell line MES-OV is an established model of ovarian endometrioid adenocarcinoma, originally derived from the malignant ascites of a patient. These adherent epithelial cells retain key characteristics of ovarian carcinoma and are widely employed in oncology research, including studies of tumor metabolism, drug response, and signal transduction. The MES-OV background is particularly suited for examining metabolic adaptations because ovarian cancer cells often exhibit altered lipid utilization compared to normal tissues.
EHHADH encodes a peroxisomal bifunctional enzyme possessing enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activities, catalyzing the second and third steps of fatty acid ??-oxidation within peroxisomes. Its expression is transcriptionally regulated by PPAR?? and activated by fatty acids and peroxisomal proliferators, while its enzymatic function generates acetyl-CoA and shortened fatty acyl-CoAs that subsequently enter mitochondrial ??-oxidation and the TCA cycle. EHHADH interacts with the peroxisomal import receptor PEX5 and collaborates with other peroxisomal enzymes, including ACOX1, HSD17B4, and SCP2, to maintain lipid homeostasis. Disruption of EHHADH therefore impedes peroxisomal ??-oxidation, potentially altering cellular energy metabolism and lipid mediator production.
In the context of ovarian endometrioid adenocarcinoma, metabolic reprogramming often involves altered fatty acid oxidation pathways, and peroxisomal dysfunction may contribute to tumor cell survival, proliferation, and drug resistance. Loss of EHHADH disrupts a key node in peroxisomal metabolism, providing a valuable tool to dissect how ovarian cancer cells adapt to impaired lipid catabolism. This polyclonal knockout model can help identify metabolic vulnerabilities that arise when peroxisomal ??-oxidation is compromised, potentially revealing new therapeutic targets.
Researchers can employ this EHHADH knockout model in a variety of experimental settings, including ??-oxidation flux measurements using labeled fatty acids, peroxisomal immunofluorescence staining, acyl-carnitine profiling by mass spectrometry, western blot detection of EHHADH, and RT-qPCR analysis of PPAR?? target genes. Additionally, functional assays assessing cell proliferation, lipid accumulation, and metabolic flux provide insight into cancer cell adaptation. These polyclonal cells are suitable for transient and stable manipulation, enabling advanced mechanistic studies of peroxisomal biology and metabolic network rewiring in ovarian cancer. For further information, please contact Ascent Research.
