The HDGF Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the heparin-binding growth factor HDGF. This product derives from the NCI-H1975 lung adenocarcinoma cell line and provides a genetically disrupted HDGF locus, resulting in a polyclonal pool of cells with heterogeneous editing events. The knockout model enables investigation of HDGF??s role in oncogenic signaling, proliferation, and drug resistance mechanisms without requiring single-cell clonal isolation.
The parental NCI-H1975 cell line is an adherent epithelial cell line isolated from a female patient with metastatic lung adenocarcinoma. It harbors the clinically relevant EGFR L858R/T790M double mutation, which confers sensitivity and acquired resistance to first- and second-generation tyrosine kinase inhibitors. This well-characterized non-small cell lung cancer (NSCLC) model exhibits rapid growth and serves as a standard platform for studying EGFR-driven tumorigenesis and therapeutic resistance.
HDGF functions both as a secreted growth factor and an intracellular transcriptional repressor. Upon secretion, it binds to cell surface nucleolin, triggering activation of the PI3K/AKT and MAPK/ERK signaling cascades. Downstream, HDGF promotes expression of Cyclin D1, VEGF, MMP-2, and MMP-9, which collectively drive cell cycle progression, angiogenesis, and matrix remodeling. Intracellularly, HDGF can repress gene transcription, interacting with factors such as SUMO1 and HDGF-related proteins. Upstream regulators include ??-catenin/TCF, STAT3, NF-??B, and hypoxia, linking HDGF expression to key oncogenic and stress response pathways.
In the context of the NCI-H1975 EGFR-mutant background, HDGF overactivity may contribute to bypass signaling that sustains proliferation and survival during EGFR inhibition. HDGF-mediated secretion of VEGF and activation of AKT and ERK1/2 are potential mediators of resistance to EGFR tyrosine kinase inhibitors. Thus, this polyclonal knockout model provides a valuable tool to dissect the HDGF-dependent signaling network in NSCLC, including crosstalk between HDGF-driven pathways and EGFR signaling.
This polyclonal HDGF knockout cell population is suitable for a range of functional assays, including monitoring changes in AKT and ERK phosphorylation by western blot, quantifying expression of target genes like Cyclin D1 and VEGF by RT-qPCR, and assessing cell proliferation using MTT assays. Migration and invasion capacity can be evaluated in Transwell systems, while apoptosis responses are measurable by Annexin V/PI staining. Immunofluorescence labeling allows visualization of HDGF, nucleolin, or downstream effectors. For additional information, please contact Ascent Research.
