The HK1 Knockout NCI-H1975 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of NCI-H1975 cells carrying a disruption of the human hexokinase-1 (HK1) gene. This polyclonal knockout cell pool provides a heterogeneous loss-of-function model for studying HK1 function in a lung adenocarcinoma background. As a polyclonal product, it captures a range of editing events across the cell population, enabling robust functional studies without the bias of single-cell cloning.
The NCI-H1975 cell line is a widely used model of human lung adenocarcinoma, derived from a female patient. It harbors EGFR L858R and T790M mutations, conferring sensitivity to first-generation EGFR inhibitors and mediating acquired resistance. These cells are extensively used to study oncogenic signaling, drug resistance, and metabolic reprogramming in non-small cell lung cancer (NSCLC).
Hexokinase-1 (HK1) catalyzes the phosphorylation of glucose to glucose-6-phosphate, the initial rate-limiting step of glycolysis, and also channels glucose into the pentose phosphate pathway. HK1 activity is upregulated by insulin, hypoxia-inducible factor 1-alpha (HIF1A), and the PI3K/Akt signaling cascade, while it is negatively regulated by AMPK under energy stress. A key feature of HK1 is its mitochondrial localization, where it binds to the voltage-dependent anion channel (VDAC). This interaction facilitates coupling of glycolysis to oxidative phosphorylation and, through association with BAD, prevents cytochrome c release and opening of the mitochondrial permeability transition pore, thereby inhibiting apoptosis. Consequently, HK1 integrates metabolic flux with cell survival decisions.
In NCI-H1975 cells, which rely on enhanced glycolysis driven by oncogenic EGFR signaling, disruption of HK1 is predicted to markedly reduce glucose phosphorylation, diminishing glycolytic intermediates and ATP synthesis. Loss of mitochondrial HK1?CVDAC binding may release pro-apoptotic constraints, sensitizing these NSCLC cells to apoptosis. This model thus provides a platform to dissect metabolic dependencies and apoptosis regulation in an EGFR-mutant lung cancer context.
Key research applications include investigating HK1’s role in glycolysis and mitochondrial apoptosis using functional assays such as glucose uptake, lactate production, ATP measurement, and Seahorse metabolic flux analysis. The effect of HK1 loss on drug sensitivity can be assessed via MTT viability and Annexin V apoptosis assays, while co-immunoprecipitation and Western blotting can probe HK1?CVDAC interactions. This polyclonal knockout population serves as a versatile tool for cancer metabolism and drug resistance studies. For further details, please contact Ascent Research.
