The CCM2 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the NCI-H1975 human lung adenocarcinoma cell line. This product provides a heterogeneous pool of edited cells with targeted disruption of the CCM2 gene, enabling loss-of-function studies without clonal selection. The polyclonal format preserves population-level diversity, which can be advantageous for examining bulk cellular responses and minimizing clone-specific artifacts. These knockout cells are designed for researchers investigating CCM2 function in a non-endothelial malignant epithelial context.
The parental NCI-H1975 cell line is a well-characterized non-small cell lung cancer (NSCLC) model established from the pleural effusion of a female patient with adenocarcinoma. These cells harbor activating mutations in the epidermal growth factor receptor (EGFR), specifically L858R in exon 21 and T790M in exon 20, resulting in constitutively active EGFR signaling and acquired resistance to first-generation tyrosine kinase inhibitors (TKIs). NCI-H1975 cells are widely employed to study EGFR-driven oncogenesis and to evaluate next-generation EGFR-targeted therapies. The epithelial origin and adherent growth properties make this line suitable for a variety of biochemical, imaging, and functional assays.
CCM2 (cerebral cavernous malformations 2) encodes a scaffold protein that nucleates the heterotrimeric CCM complex by interacting with KRIT1 (CCM1) and PDCD10 (CCM3). Through this complex, CCM2 negatively regulates the MEKK3-MEK5-ERK5 kinase cascade and RhoA-ROCK signaling to maintain cell?Ccell junction integrity. Upstream, CCM2 expression is regulated by Notch1 intracellular domain (NICD1) and HES1, and it is modulated by hemodynamic shear stress and ICAP1 (ITGB1BP1). Downstream targets include the ERK5-dependent transcription factors KLF2 and KLF4 and RhoA effectors ROCK1/2. Loss of CCM2 disrupts these inhibitory mechanisms, resulting in hyperactivation of ERK5 and RhoA, destabilization of VE-cadherin junctions, and a pro-angiogenic phenotype characteristic of cerebral cavernous malformations.
Although CCM2 is predominantly studied in endothelial cells, its knockout in the NCI-H1975 adenocarcinoma background offers a unique opportunity to dissect CCM2 function in a malignant epithelial setting. The NCI-H1975 line??s constitutive EGFR signaling provides a platform to explore potential crosstalk between CCM2-mediated pathways and oncogenic kinase cascades. For instance, EGFR-driven MEK5-ERK5 and RhoA activation may exacerbate or compensate for CCM2 loss, thereby revealing context-dependent roles in tumor cell adhesion, migration, and invasion. This model is particularly relevant for investigating whether CCM2 deficiency in non-endothelial cells contributes to tumor progression or alters therapeutic sensitivity.
This polyclonal knockout pool is suitable for transwell migration and invasion assays, RhoA-GTP pull-down, phospho-ERK5 immunoblotting, and immunofluorescence analysis of F-actin and VE-cadherin. Proliferation can be measured by MTT assay, and in vivo tumorigenicity assessed by xenograft models. Additionally, the cells can be applied to drug screening for RhoA/ROCK or ERK5 inhibitors in a non-endothelial context. For detailed technical specifications and to discuss your specific research needs, please contact Ascent Research.
