HPCAL1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the HPCAL1 gene has been disrupted via CRISPR/Cas9-mediated gene editing. The product consists of a heterogeneous pool of NCI-H1975 cells harboring diverse loss-of-function mutations in HPCAL1, providing a robust model to study gene function without clonal selection artifacts. This knockout system enables investigation of HPCAL1-dependent processes in a human lung adenocarcinoma background, serving as a versatile tool for cancer biology and signal transduction research.
The host cell line NCI-H1975 is a KRAS-mutant, TP53-mutant human lung adenocarcinoma epithelial line derived from a metastatic pleural effusion, representing an aggressive non-small cell lung carcinoma (NSCLC) model. These genetic lesions mimic common driver mutations found in advanced lung cancer, rendering the cells highly tumorigenic and metastatic. NCI-H1975 exhibits typical adenocarcinoma morphology and growth characteristics, and its well-characterized signaling profile makes it a standard platform for NSCLC research. Retention of the KRAS/TP53-mutant background in the knockout cells allows direct dissection of HPCAL1 contributions to malignant phenotypes.
HPCAL1 (hippocalcin-like 1) is a neuronal calcium sensor that modulates GPCR signaling by directly interacting with GRK5 and beta-arrestin-2. Upon intracellular Ca2? elevation, HPCAL1 facilitates GPCR desensitization and clathrin-dependent internalization, linking calcium transients to receptor trafficking. Downstream, HPCAL1 promotes activation of the MAPK/ERK pathway through MAPK1/3 (ERK1/2), thereby regulating cell proliferation and migration. Additional interacting factors include dopamine D2 and adenosine A2A receptors, further implicating HPCAL1 in fine-tuning GPCR responsiveness. This calcium-dependent signaling hub integrates extracellular cues to control cellular behavior.
In the NCI-H1975 NSCLC context, HPCAL1 overexpression is associated with enhanced proliferation and motility, potentially via sustained MAPK/ERK signaling. The knockout model ablates HPCAL1 function, enabling researchers to assess its role in tumor cell migration, invasion, and GPCR signaling dynamics. Studying HPCAL1 disruption in a KRAS/TP53-mutant background is particularly valuable for elucidating crosstalk between oncogenic drivers and calcium sensor proteins. This loss-of-function system can reveal HPCAL1-dependent pathways contributing to lung adenocarcinoma progression and metastasis, aiding in the identification of therapeutic targets.
Typical experimental applications include confirmation of knockout via western blotting and RT-qPCR, immunofluorescence to assess protein localization, and functional assays such as MTS proliferation, wound healing migration, and Boyden chamber invasion. GPCR internalization assays and calcium imaging permit quantitative analysis of receptor trafficking and calcium dynamics. MAPK phosphorylation profiling evaluates downstream pathway activity. These applications support drug target validation, GPCR pharmacology, and metastasis studies. For detailed protocols, pricing, or custom project inquiries, please contact Ascent Research.
