The HSD17B7 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, designed for loss-of-function studies of the HSD17B7 gene. This heterogeneous pool is generated through CRISPR/Cas9-mediated gene disruption, providing a versatile model to investigate HSD17B7-dependent steroid hormone metabolism and cholesterol biosynthesis pathways. The polyclonal nature preserves genetic diversity, enabling robust analysis of gene function in a physiologically relevant tumor microenvironment context.
HCT 116 cells serve as a well-characterized model for colorectal cancer research, originating from a human male colorectal carcinoma with microsatellite instability. They harbor an oncogenic KRAS G13D mutation and a stabilizing CTNNB1 mutation that leads to constitutive Wnt/??-catenin signaling, contributing to their tumorigenic properties. These genetic features make HCT 116 cells particularly suited for exploring the intersection of oncogenic signaling with steroid metabolism.
HSD17B7 encodes 17??-hydroxysteroid dehydrogenase type 7, an enzyme that catalyzes the final step in estradiol biosynthesis from estrone and also converts androstenedione to testosterone, thereby fueling estrogen and androgen signaling. In addition, HSD17B7 plays a role in post-squalene cholesterol biosynthesis by reducing zymosterol, linking steroidogenesis directly to cellular cholesterol metabolism. The gene is transcriptionally regulated by SF-1 (NR5A1) and SREBP2, and its activity is integrated with estrogen receptor alpha (ESR1) signaling. HSD17B7-generated estradiol and testosterone act downstream through ESR1 and the androgen receptor (AR), respectively, while its role in cholesterol synthesis involves interactions with cytochrome P450 enzymes, 3??-hydroxysteroid dehydrogenase, and other short-chain dehydrogenase/reductase (SDR) family members. The broader metabolic network includes CYP19A1, STS, HSD3B2, SOAT1, and CYP51A1, positioning HSD17B7 at a critical node connecting steroid hormone production and cholesterol homeostasis.
In HCT 116 colorectal cancer cells, disruption of HSD17B7 is expected to impair local estradiol and testosterone synthesis, attenuating estrogen receptor and androgen receptor activity, while concurrently altering cholesterol biosynthesis through zymosterol accumulation. Given that HCT 116 cells possess activated Wnt/??-catenin and MAPK pathways due to CTNNB1 and KRAS mutations, the intersection between steroid signaling and oncogenic growth pathways can be systematically dissected. This polyclonal knockout model thus provides a powerful platform to elucidate how HSD17B7-dependent steroidogenesis influences proliferation, membrane composition, and therapy resistance in a colorectal carcinoma background.
Researchers can employ these polyclonal knockout cells in a broad array of applications, including quantification of intracellular estradiol and testosterone via ELISA, profiling of cholesterol intermediates through chromatographic methods, and transcriptomic analysis by RNA-seq to map HSD17B7-dependent gene networks. Comparative Western blotting and RT-qPCR enable verification of target disruption and assessment of compensatory pathway activation, while cell proliferation assays reveal functional consequences in colorectal cancer contexts. Further, these cells are amenable to drug sensitivity screens to interrogate the role of steroid metabolism in chemoresistance. For additional details or technical inquiries, please contact Ascent Research.
