Pten Knockout LLC Cell Line

The Pten Knockout LLC Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the Lewis lung carcinoma (LLC) cell line. The product provides a targeted loss-of-function model for the phosphatase and tensin homolog (Pten) gene in a murine lung cancer background. Through CRISPR/Cas9-mediated gene disruption, the Pten locus is inactivated, enabling researchers to study the downstream consequences of PTEN deficiency in a genetically defined system. This stable knockout cell line is suitable for a broad range of functional assays and is supplied as a ready-to-use culture for immediate experimental application.

The LLC cell line is a widely used model of non-small cell lung carcinoma originally established from a spontaneous tumor in a C57BL/6 mouse. It exhibits aggressive growth characteristics in vitro and in vivo, forming tumors efficiently in syngeneic immunocompetent hosts. The C57BL/6 background provides a defined genetic context that is particularly useful for tumor immunology studies, as it allows engraftment into wild-type C57BL/6 mice without immune rejection. The Lewis lung carcinoma model has been extensively employed for metastasis research, angiogenesis studies, and preclinical drug testing.

PTEN functions as a lipid phosphatase that dephosphorylates phosphatidylinositol (3,4,5)-trisphosphate (PIP3) to produce phosphatidylinositol (4,5)-bisphosphate (PIP2), directly opposing phosphatidylinositol 3-kinase (PI3K) activity. This enzymatic activity serves as a critical negative regulator of AKT signaling: by reducing PIP3 levels, PTEN impairs PDK1-mediated AKT recruitment to the plasma membrane and subsequent phosphorylation. Consequently, PTEN inactivation leads to constitutive AKT activation, which in turn phosphorylates and inhibits downstream substrates such as GSK3?? and FOXO transcription factors, while activating mTORC1. The Pten gene is transcriptionally regulated by p53 and NF-??B and is post-transcriptionally modulated by microRNAs including miR-21 and miR-214. PTEN is stabilized at the membrane through interactions with scaffolding proteins such as MAGI and NHERF, ensuring proper subcellular localization and function.

Loss of PTEN function is frequently observed in lung cancer and has been associated with enhanced tumor cell proliferation, survival, migration, and resistance to therapy. In the LLC model, Pten knockout enables dissection of PTEN-dependent tumor-suppressive mechanisms in a cell-autonomous manner. The combination of PTEN deficiency and the highly tumorigenic LLC background creates a powerful system for evaluating the role of hyperactivated PI3K/AKT/mTOR signaling in driving aggressive lung cancer phenotypes. This model is particularly valuable for studying the interplay between PTEN status and other common oncogenic alterations found in lung adenocarcinoma and squamous cell carcinoma.

Researchers can employ the Pten Knockout LLC Cell Line to investigate PTEN tumor suppressor function in lung cancer, including assessment of AKT pathway activation via phospho-AKT western blotting, cell proliferation using MTT or BrdU incorporation assays, and apoptosis detection through cleaved caspase-3 measurement. Migratory and invasive potential can be examined using Boyden chamber assays, while anchorage-independent growth is assessed by soft agar colony formation. For in vivo studies, these cells reliably form tumors in C57BL/6 syngeneic models, facilitating xenograft experiments to test PI3K/AKT/mTOR inhibitors such as rapamycin or dual PI3K/mTOR antagonists. The model also supports metastasis research and evaluation of combination therapies targeting PTEN-deficient tumors. For further information or to place an order, please contact Ascent Research.