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The A431 cell line, derived from a solid epidermoid carcinoma tumor in an 85-year-old female patient, is a human tumor cell line with an epithelial morphology, typically growing in clusters. The A-431 cell line is extensively utilized in cancer, toxicity, and immuno-oncology studies, serving as a positive control for epidermal growth factor (EGF) receptor expression due to its high receptor density.
Upon binding of EGF to its receptor (EGFR) on the surface of A431 cells, a rapid tyrosine phosphorylation of membrane proteins occurs, triggering a cascade of intracellular signaling pathways. These pathways include the MAPK/ERK and PI3K/AKT pathways, which are pivotal in regulating cell cycle progression, survival, and proliferation.
EGFR stimulates cell proliferation at low concentrations, whereas at higher concentrations, it inhibits growth and induces terminal differentiation in A431 cells. This dynamic response to EGFR underscores the cell line's utility in exploring cell signaling pathways and the cell cycle in the context of cancer.
A-431 cell-derived xenograft models are used for studying tumor behavior in a live environment and evaluating anticancer therapies. These models help assess how treatments such as EGF supplementation and radiation, affect tumor growth and highlight the cells' sensitivity to radiation.
In summary, the A-431 cell line serves as an invaluable human epidermoid carcinoma cell model, facilitating a deeper understanding of EGFR signaling, tumor biology, and the development of therapeutic interventions aimed at combating epidermoid carcinoma and other related cancers.
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- Receptors expressed: EGF-binding sites
- Protein expression: p53 positive
- Isoenzymes: G6PD, B, PGM1, 1, PGM3, 1, ES-D, 1, Me-2, 0, AK-1, 1, GLO-1, 2
- Tumorigenic: Yes, in immunosuppressed mice
- Products: HBp17
- Mutational profile: BRAF V600Ewt
- Karyotype: Six marker chromosomes with rearrangements: der(6), der(7), der(17), der(21), dic(13,14), and dic(14,18). Amplification of the C-MYC oncogene at 8q24 in two marker chromosomes: dup(8)(q24) and der(15)t(8,15)(q22,p11).
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- 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 Accutase, 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.
- seedingDensity: 1 x 104 cells/cm2 will result in a confluent monolayer within 4 days.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- 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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