The KHK Knockout HCT 116 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population from the HCT 116 human colorectal carcinoma line, with targeted disruption of the KHK gene. This heterogeneous pool of loss-of-function mutants serves as a robust model for investigating fructose metabolism, avoiding clonal artifacts and enabling parallel experimental replicates.
The HCT 116 cell line is a widely utilized model of colorectal carcinoma, featuring microsatellite instability (MSI-H) and a mutant KRAS allele (G13D). These characteristics drive oncogenic MAPK signaling and metabolic reprogramming, making the line highly relevant for cancer metabolism studies and therapeutic testing.
KHK encodes ketohexokinase, which catalyzes the phosphorylation of fructose to fructose-1-phosphate, the entry step into glycolysis and lipogenesis. KHK activity is regulated by upstream cues including ChREBP, insulin, glucagon, and HIF-1??, linking fructose metabolism to hormonal and hypoxic signals. Fructose-1-phosphate feeds into glycolytic intermediates, promotes lipogenesis, and modulates downstream effectors mTOR and AMPK, thereby coordinating nutrient sensing with cellular growth. KHK interacts with aldolase B, and along with triose phosphate isomerase, constitutes a core metabolic node. In KRAS-mutant colorectal cancer cells, this pathway enables alternative substrate utilization, sustaining biosynthesis and proliferation under fluctuating nutrient conditions.
Knockout of KHK in HCT 116 cells disrupts fructose-dependent metabolic pathways, offering a platform to examine how loss of fructose catabolism impacts colorectal cancer cell proliferation and survival, particularly in the context of mutant KRAS-driven metabolic stress. This model aids in dissecting the interplay between fructose metabolism and key signaling hubs such as mTOR and AMPK.
This polyclonal knockout cell population is ideal for metabolic flux analyses (Seahorse), proliferation (MTT, BrdU) and colony formation assays, and transcriptomic profiling by RNA-seq or RT-qPCR. Western blotting confirms KHK ablation and downstream signaling alterations. Typical research applications include functional genomics of fructose-dependent cancer metabolism, drug target validation for metabolic syndrome and NAFLD, and mechanistic studies of nutrient?Concogene interactions in KRAS-driven tumors. For additional information on experimental optimization, please contact Ascent Research.
