HSD17B11 Knockout HT29 Polyclonal Cells comprise a heterogeneous population of CRISPR/Cas9-edited HT29 human colorectal adenocarcinoma cells, designed to disrupt the HSD17B11 locus and create a loss-of-function model for studying this short-chain dehydrogenase/reductase family member. The polyclonal format preserves genetic diversity while enabling robust functional screening, offering a flexible tool for investigating HSD17B11-dependent processes in epithelial cancer biology, particularly those related to steroid metabolism and retinoid signaling.
The HT29 cell line is an established human colorectal adenocarcinoma model originally derived from a primary tumor of a 44-year-old female patient. These cells exhibit characteristic epithelial morphology and retain key features of intestinal differentiation, making them a widely used platform for dissecting colorectal cancer pathogenesis, intestinal epithelial function, and hormone-responsive signaling pathways.
HSD17B11 encodes an oxidoreductase that catalyzes NAD+/NADH-dependent interconversions critical to steroid and retinoid metabolism. It converts 17-beta-hydroxysteroids??including testosterone and estradiol??to their less active 17-keto derivatives, thereby modulating androgen and estrogen receptor activity. Additionally, HSD17B11 oxidizes retinol in the retinoic acid synthesis pathway, influencing all-trans-retinoic acid levels and downstream retinoid X receptor/retinoic acid receptor (RXR/RAR) signaling. The enzyme interacts with cellular retinoic acid-binding proteins CRABP1 and CRABP2 and aldo-keto reductase family members, and is transcriptionally regulated by peroxisome proliferator-activated receptor gamma (PPARG), androgen receptor (AR), sterol regulatory element-binding factor 1 (SREBF1), and RAR/RXR heterodimers. Downstream effectors include androgen-responsive genes such as PSA/KLK3 and retinoic acid-responsive targets like RARB and CYP26A1, linking HSD17B11 to cell cycle control and differentiation programs.
In the context of colorectal adenocarcinoma, HSD17B11 knockout in HT29 cells provides a physiologically relevant system to interrogate the interplay between local steroid hormone metabolism and retinoid signaling. Because colorectal tissues are exposed to dietary retinoids and express steroid-converting enzymes, loss of HSD17B11 may alter the balance between growth-promoting and differentiation-inducing signals. This model is particularly pertinent to colorectal cancer research, where aberrant steroid metabolism and retinoic acid insensitivity have been implicated in tumor progression. Moreover, the polyclonal knockout population facilitates rapid assessment of HSD17B11??s role in modulating sensitivity to anti-androgens, retinoids, and metabolic stress.
Typical applications include examining the impact of HSD17B11 disruption on cell proliferation, apoptosis, and colony formation using standard MTT and Annexin V assays. Quantitative RT-PCR and western blotting enable validation of altered expression of key steroid receptors and retinoid-responsive genes, while liquid chromatography-mass spectrometry (LC-MS) can quantify changes in intracellular steroids and retinoids. This model also supports drug sensitivity screens to test inhibitors of androgen signaling, retinoic acid metabolism, or downstream pathways. By combining the HT29 background with polyclonal HSD17B11 knockout, researchers can dissect the enzyme??s contributions to colorectal cancer cell biology and evaluate its potential as a therapeutic target. For further technical specifications or assistance, please contact Ascent Research.
