The INSR Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line. This product offers a genetically heterogeneous pool of cells with disrupted INSR expression, creating a versatile tool for studying insulin receptor function and insulin signaling pathways in a cancer-relevant context.
The parental A-549 cell line originates from a human lung adenocarcinoma and retains epithelial morphology. A-549 cells are extensively characterized and widely employed in cancer research, particularly for studies of non-small cell lung cancer (NSCLC) biology, drug response, and metabolic reprogramming. Their adherent growth and well-documented signaling properties make them a suitable chassis for investigating the tumor-promoting roles of receptor tyrosine kinases such as INSR.
INSR encodes a tetrameric receptor tyrosine kinase that binds insulin, IGF-1 and IGF-2, initiating autophosphorylation and recruitment of adaptors including IRS1, SHC1 and CBL. Activated INSR propagates signals through two principal cascades: the PI3K-Akt pathway, whereby IRS1 engages PI3K to phosphorylate Akt, regulating mTOR and FOXO1, and the Ras-MAPK pathway, driven by the SHC1?CGRB2?CSOS complex, leading to ERK1/2 (MAPK1/3) phosphorylation. INSR activity also governs SREBP1-mediated lipogenesis and is counterbalanced by TNF-alpha and the phosphatase PTPN1 (PTP1B).
In lung adenocarcinoma A-549 cells, INSR signaling contributes to metabolic adaptation and proliferative capacity, making this knockout model valuable for examining insulin-dependent growth and metabolic reprogramming in cancer. Disruption of INSR in this cell line can help elucidate mechanisms of insulin resistance, a condition often associated with type 2 diabetes and obesity, and its intersection with oncogenic pathways. Moreover, the polyclonal nature of the population may reveal heterogeneous responses to insulin pathway inhibitors and aid in identifying resistance mechanisms.
Researchers can utilize these INSR knockout A-549 polyclonal cells in a variety of assays, including western blot analysis of phosphorylated Akt (Ser473) and ERK (Thr202/Tyr204) following insulin stimulation, RT-qPCR quantitation of metabolic gene expression, and flow-cytometric measurement of glucose uptake using fluorescent glucose analogs. Proliferation and viability can be monitored via MTS/MTT assays, while wound-healing and transwell assays assess migration and invasion. Applications extend to metabolomic profiling under insulin-depleted or receptor-inhibited conditions, and the cells are compatible with high-throughput screening of insulin receptor-targeted compounds. For technical assistance and ordering details, contact Ascent Research.
