The ECI2 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 human lung adenocarcinoma cell line, harboring a targeted disruption of the ECI2 gene. This loss-of-function model eliminates the activity of mitochondrial enoyl-CoA delta isomerase 2, a critical enzyme in the ??-oxidation of unsaturated fatty acids, enabling researchers to dissect the role of fatty acid catabolism in cancer cell metabolism, mitochondrial function, and therapeutic response.
The parental NCI-H1299 cell line is a well-characterized non-small cell lung cancer model established from a lymph node metastasis of a 43-year-old male. It retains wild-type p53 and is widely employed to investigate apoptosis, invasion, metastatic potential, and drug sensitivity, particularly in the context of non-small cell lung carcinoma. This cellular background provides a physiologically relevant setting to assess the consequences of impaired unsaturated fatty acid degradation.
ECI2 catalyzes the isomerization of 3-cis-??6-enoyl-CoA to 2-trans-??6-enoyl-CoA, a requisite step for mitochondrial ??-oxidation of unsaturated fatty acids. Its expression is regulated by PPAR??, PPAR??, and PGC-1??, and is responsive to SIRT1 and AMPK. Downstream, ECI2 supports acetyl-CoA and NADH production for ATP synthesis and ketone body generation. It interacts with enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and the mitochondrial trifunctional protein (HADHA/HADHB), functioning alongside SCP2, ACADVL, and ACAA2 in the fatty acid ??-oxidation pathway.
In the NCI-H1299 background, ECI2 disruption is expected to compromise the utilization of unsaturated fatty acids, forcing a metabolic shift that may reveal dependencies on alternative lipid or glucose substrates. This model is particularly relevant for non-small cell lung cancer, where altered fatty acid oxidation has been linked to tumor progression and drug resistance. By abrogating ECI2 function, researchers can explore how mitochondrial lipid handling influences cell proliferation, apoptosis, and invasive capacity, and whether targeting this node sensitizes cancer cells to existing therapies.
Typical applications include metabolic flux analysis using 3H-palmitate, Seahorse-based mitochondrial respiration assays, RT-qPCR and immunoblotting for pathway validation, and cell viability assessments under lipid-rich conditions. The polyclonal population is also amenable to apoptosis and invasion assays, and studies of mitochondrial membrane potential. This model is a robust tool for examining lipid metabolism in lung cancer. For technical support, please contact Ascent Research.
