In Stock Cell Lines
Homo sapiens (Human)
Lung
Adherent
The PTEN Knockout A549 Cell Line provides a CRISPR/Cas9-edited loss-of-function model in A549 lung adenocarcinoma cells. By eliminating PTEN, a lipid phosphatase that opposes PI3K/AKT signaling, this cell line exhibits constitutive AKT activation and deregulated mTORC1, FOXO, and GSK3?? pathways. Applications include evaluating PI3K/AKT/mTOR inhibitors, studying migration and apoptosis, and investigating tumor suppressor cooperation in a KRAS-mutant, p53 wild-type background. Representative assays include phospho-AKT Western blot and wound healing migration assays.
The PTEN Knockout A549 Cell Line is a CRISPR/Cas9-edited knockout cell line disrupting PTEN expression in the A549 human lung adenocarcinoma epithelial background. This loss-of-function model enables investigation of PTEN tumor suppressor functions and PI3K/AKT/mTOR pathway dysregulation in a defined oncogenic context. The cell line provides a robust tool for studying signaling mechanisms and evaluating targeted therapies.
The A549 cell line, isolated from human lung carcinoma tissue, exhibits epithelial morphology and harbors a KRAS G12S mutation with wild-type p53. It serves as a model for type II alveolar epithelial cells and is widely used in non-small cell lung adenocarcinoma research. The KRAS mutation creates a genetic setting in which PTEN loss amplifies downstream signaling, facilitating studies of oncogenic cooperation.
PTEN is a dual-specificity phosphatase that primarily dephosphorylates PIP3 to PIP2, opposing PI3K-mediated AKT activation. It also possesses protein phosphatase activity implicated in migration and genomic stability. Upstream regulators include the transcription factor p53 and microRNA miR-21, while protein kinases such as CSNK2A1 and GSK3B modulate PTEN activity. Downstream, PTEN loss leads to constitutive AKT1 activation, promoting mTORC1 signaling, GSK3?? inhibition, and phosphorylation of FOXO1/3, BAD, and MDM2. This results in enhanced cell survival, proliferation, and migration, while relieving cell cycle brakes via CDKN1A and CDKN1B downregulation. PTEN interacts with scaffolding proteins MAGI1-3 and is targeted for degradation by NEDD4L, further diversifying its regulatory network.
In A549 cells, PTEN knockout synergizes with the KRAS G12S mutation to hyperactivate PI3K/AKT/mTOR signaling, mimicking aggressive lung adenocarcinoma genotypes. This model enables dissection of crosstalk between RAS-ERK and PI3K pathways and investigation of resistance mechanisms to RAS-directed therapies. The p53 wild-type background permits analysis of PTEN-p53 interactions in apoptosis and senescence, offering a versatile platform for studying tumor suppressor cooperation and identifying synthetic lethal targets.
Applications include testing PI3K/AKT/mTOR inhibitors (e.g., LY294002, rapamycin) via proliferation (MTS) and colony formation assays. Signaling readouts such as phospho-AKT (Ser473) Western blot, phospho-kinase arrays, and RT-qPCR for PTEN transcript can be used to confirm pathway modulation. Cell migration and invasion are assessable by wound healing and Transwell, while flow cytometry (Annexin V) measures apoptosis. Xenograft models permit in vivo tumorigenicity and drug response evaluation. For further details, please contact Ascent Research.
