The ING1 Knockout NCI-H1975 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout population derived from the human lung adenocarcinoma cell line NCI-H1975. CRISPR/Cas9-mediated gene disruption has been used to target the ING1 tumor suppressor, generating a heterogeneous cell pool with loss-of-function mutations. This polyclonal format avoids clonal selection bias and provides a robust model for studying ING1-dependent signaling in non-small cell lung cancer (NSCLC).
The parental NCI-H1975 line originates from a female never-smoker with lung adenocarcinoma and harbors an activating EGFR L858R mutation, which drives constitutive oncogenic signaling. The cells exhibit moderate EGFR expression and sensitivity to EGFR tyrosine kinase inhibitors. Combined with ING1 knockout, this background enables dissection of cooperative effects between EGFR-driven proliferation and loss of tumor suppression.
ING1 is a type II tumor suppressor that modulates cell cycle arrest, apoptosis, and senescence via p53-dependent and -independent mechanisms. It physically interacts with histone acetyltransferases EP300 and CREBBP, as well as HDAC1?CSIN3A?CSAP30 corepressor complexes, to regulate histone acetylation and chromatin structure. ING1 binds TP53 and enhances p53 acetylation, promoting transcription of target genes including CDKN1A (p21), BAX, and BBC3 (PUMA). It also associates with PCNA to couple DNA replication to DNA damage responses. Upstream signals such as DNA damage, oxidative stress, and TGF-beta stabilize ING1, potentiating p53-mediated growth suppression.
In NCI-H1975 cells, ING1 disruption impairs p53 acetylation and transcriptional activity, resulting in reduced expression of p21, BAX, and PUMA. This leads to diminished apoptosis and senescence in response to genotoxic stress, while oncogenic EGFR L858R maintains proliferative signaling. The model thus replicates a two-hit scenario relevant to NSCLC pathogenesis and is well-suited for studying the intersection of EGFR and p53 pathways.
These cells support diverse applications including Western blotting for ING1, p53, acetyl-p53, and p21; RT-qPCR analysis of downstream targets; and functional assays such as MTT viability, Annexin V apoptosis, and cell cycle flow cytometry. They facilitate drug sensitivity testing for erlotinib and SAHA, senescence ??-galactosidase assays, and ChIP for histone acetylation. The model is ideal for tumor suppressor research, EGFR inhibitor studies, and HDAC inhibitor screening. For additional information, please contact Ascent Research.
