The ITPRIPL2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population featuring targeted disruption of ITPRIPL2 in the human NCI-H1975 lung adenocarcinoma cell line. This loss-of-function model enables investigation of ITPRIPL2, a putative regulator of IP3 receptor (IP3R) activity, in non-small cell lung cancer (NSCLC). The polyclonal pool preserves genetic heterogeneity, providing a robust platform for studying calcium signaling dynamics and cancer cell behavior.
The NCI-H1975 cell line, derived from a non-smoking female with NSCLC, harbors activating EGFR mutations (L858R and T790M) and wild-type KRAS. These genetic features drive constitutive EGFR signaling and resistance to first-generation tyrosine kinase inhibitors (TKIs), establishing NCI-H1975 as a key model for acquired drug resistance and metastasis in lung adenocarcinoma. The T790M gatekeeper mutation is particularly relevant for studying next-generation EGFR inhibitors.
ITPRIPL2 is believed to modulate IP3R-mediated endoplasmic reticulum calcium release by interacting with ITPR1, ITPR2, and ITPR3. Downstream, calcium fluxes activate calmodulin and calcineurin, leading to NFAT dephosphorylation and nuclear translocation. This transcriptionally regulates apoptosis effectors (Bcl-2 family, caspases) and cell cycle genes. Knockout of ITPRIPL2 is expected to disrupt this signaling axis, altering calcium oscillation patterns and potentially rewiring apoptotic and proliferative programs.
In the NCI-H1975 background, oncogenic EGFR signaling couples to calcium pathways via PLC?? and IP3 generation. ITPRIPL2 loss may therefore recalibrate calcium-dependent feedback loops that influence sensitivity to EGFR TKIs. Disrupted IP3R regulation could enhance or suppress pro-apoptotic calcium signals induced by targeted therapies, affecting cell survival, migration, and invasion. This makes the model valuable for dissecting how calcium homeostasis intersects with EGFR-driven oncogenesis and drug resistance.
Applications include calcium imaging (Fluo-4 AM), Western blotting for IP3R-effector cascades, apoptosis (Annexin V/PI) and proliferation (MTS) assays, migration studies, and drug sensitivity profiling with EGFR inhibitors (e.g., osimertinib). RT-qPCR of calcium-related genes complements phenotypic analyses. Typical research uses encompass calcium signaling studies in NSCLC, functional genomics of ITPRIPL2, and drug target validation. For further details, contact Ascent Research.
