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The NCI-H2122 cell line is a human non-small cell lung cancer (NSCLC) model derived from an adenocarcinoma patient. It is notable for its harboring of a KRAS G12C mutation, a hallmark of NSCLC that leads to constitutive activation of the MAPK signaling pathway. This cell line is extensively used in studies focusing on therapeutic interventions targeting KRAS G12C and associated downstream pathways, particularly those involving MEK and ERK inhibitors. Research utilizing NCI-H2122 has highlighted its role in understanding drug resistance mechanisms and in optimizing combination therapies.
Preclinical studies using the NCI-H2122 cell line have demonstrated its utility in exploring resistance to MAPK pathway inhibitors. For instance, CRISPR screening approaches have identified MAPK7 (ERK5) as a critical mediator of pathway reactivation following MEK inhibition, suggesting potential combination strategies using MEK inhibitors like cobimetinib and MAPK7 inhibitors. The line also serves as a model for evaluating the efficacy of small molecule inhibitors, including those targeting PI3K and BRAF, which are relevant in combination with KRAS-specific treatments.
NCI-H2122 is also employed in investigating metabolic vulnerabilities in NSCLC. Studies have implicated serine biosynthesis and the folate cycle as metabolic pathways contributing to resistance against targeted therapies, such as BRAF inhibitors. Metabolic modulators like methotrexate and serine-deprivation strategies have been tested on this cell line, providing insights into overcoming drug resistance and identifying new metabolic targets for therapeutic exploitation.
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Mutational profile: Mutation: KRAS, p.Gly12Cys (c.34G>T), homozygous; Mutation: TP53, p.Gln16Leu (c.47A>T), heterozygous; Mutation: TP53, p.Cys176Phe (c.527G>T), heterozygous
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with TrypLE Express, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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