The HSDL2 Knockout A-549 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population derived from the A-549 human lung adenocarcinoma cell line, designed for the disruption of the HSDL2 gene. This polyclonal knockout cell pool provides a loss-of-function model for investigating HSDL2??s role in peroxisomal fatty acid metabolism and oncogenic signaling. The cells are generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous population with targeted inactivation of HSDL2, suitable for functional studies without clonal selection bottlenecks.
The host A-549 cell line is a widely used in vitro model of human lung adenocarcinoma, established from the tumor tissue of a 58-year-old male. These cells exhibit characteristics of type II alveolar epithelial cells and are frequently employed to study non-small cell lung cancer biology, including proliferation, metastasis, and drug resistance. A-549 cells harbor wild-type TP53 and express EGFR, making them a relevant context for exploring signaling pathways implicated in pulmonary carcinogenesis.
HSDL2 encodes a peroxisomal enzyme involved in the ??-oxidation of fatty acids. At the molecular level, HSDL2 promotes cancer cell growth and invasion primarily through activation of the AKT/mTOR signaling axis. Upstream, HSDL2 is regulated by miR-195-5p and EGF/EGFR signaling. Downstream, it activates AKT, which in turn phosphorylates mTOR, S6K, and 4E-BP1, leading to increased protein synthesis and cell cycle progression via upregulation of Cyclin D1 and CDK4. Additionally, HSDL2 enhances the expression of anti-apoptotic Bcl-2 while suppressing pro-apoptotic Bax, and it drives glycolysis by upregulating HK2 and LDHA. HSDL2 interacts with PEX14, a peroxisomal membrane protein, linking its metabolic functions to organelle biology.
In the A-549 lung adenocarcinoma model, HSDL2 knockout disrupts peroxisomal fatty acid metabolism and impairs AKT/mTOR signaling, resulting in diminished proliferative, migratory, and invasive capacities. This metabolic reprogramming is accompanied by reduced glycolytic enzyme expression and increased apoptosis. Thus, these polyclonal knockout cells offer a physiologically relevant system to dissect the crosstalk between peroxisomal lipid metabolism and oncogenic signaling, and to evaluate vulnerabilities in lung cancer cells that are dependent on HSDL2 function.
These cells are ideally suited for a range of research applications, including cancer metabolism studies, investigation of AKT/mTOR signaling dynamics, peroxisomal biology research, and drug target validation. Representative assays include western blotting for phosphorylated AKT, mTOR, and S6K; CCK-8 proliferation assays; transwell migration and invasion assays; Annexin V/PI flow cytometry for apoptosis; glycolysis stress tests using Seahorse analyzers; and fatty acid oxidation measurements. For additional technical details or to discuss your specific experimental needs, please contact Ascent Research.
