The HSD17B8 Knockout A-549 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma epithelial cell line. This loss-of-function model features targeted disruption of the HSD17B8 gene, which encodes 17-beta-hydroxysteroid dehydrogenase 8. By delivering a heterogeneous pool of knockout cells, researchers gain a genetically diverse resource to interrogate HSD17B8-dependent pathways while avoiding the selection bias inherent in monoclonal lines. The polyclonal format is especially suited for studying gene function in a population context, reflecting the cellular heterogeneity observed in tumor biology.
The A-549 host cell line was originally established from the lung adenocarcinoma of a 58-year-old male and is widely employed as a model of alveolar type II pneumocytes. These cells retain key features of pulmonary epithelial biology and are extensively utilized in in vitro oncological and pharmacological studies, particularly for non-small cell lung cancer. Their well-characterized genotype and robust growth characteristics make them an ideal chassis for exploring how hormonal and metabolic reprogramming influence lung cancer progression.
HSD17B8 catalyzes the NAD+-dependent oxidoreduction of estradiol to estrone and androstenedione to testosterone, positioning it at a critical node in steroid hormone biosynthesis. Additionally, it functions as a 3-ketoacyl-CoA reductase within the mitochondrial fatty acid elongation cycle, interacting with HSD17B4, ACAA2, HADHB, and MECR. Transcription of HSD17B8 is regulated by nuclear receptors SF-1 (NR5A1), estrogen receptor alpha (ESR1), and androgen receptor (AR), while PPAR-alpha provides metabolic control. Downstream, HSD17B8 activity modulates the expression of estrogen-responsive genes (PGR, TFF1) and androgen-responsive targets (KLK3, NKX3-1), as well as lipogenic enzymes FASN and SCD1. Thus, HSD17B8 serves as an integrator of endocrine and lipid signals.
In the A-549 lung adenocarcinoma context, disruption of HSD17B8 is predicted to upset the balance between active estradiol/testosterone and their less potent counterparts, potentially altering hormone-driven proliferation and migration. Because these cells mirror alveolar type II pneumocyte traits, the knockout model enables dissection of how steroid interconversion and mitochondrial fatty acid synthesis converge to shape tumor metabolism and malignant behavior. This makes the model relevant for studies of hormone-dependent lung cancers, metabolic reprogramming in tumors, and disorders such as polycystic ovary syndrome where these pathways are dysregulated.
This polyclonal knockout population supports a broad range of experimental applications. Researchers can employ RT-qPCR and Western blotting to validate HSD17B8 loss and assess compensatory pathways. LC-MS-based steroid profiling permits quantitative tracking of estradiol, estrone, testosterone, and androstenedione, while RNA-seq illuminates transcriptomic changes. Functional assays including proliferation, migration/invasion, and fatty acid oxidation assays help elucidate the gene’s impact on cancer cell fitness and lipid utilization. Hormone-responsive element reporter assays and co-immunoprecipitation can further probe interactions with ESR1 and AR. Additionally, drug sensitivity testing against steroid pathway inhibitors may guide therapeutic strategies. For further assistance, please contact Ascent Research.
