The HECA Knockout NCI-H1975 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the HECA gene in the NCI-H1975 human lung adenocarcinoma epithelial cell line. This loss-of-function model enables dissection of HECA-mediated transcriptional and post-transcriptional regulation in a lung cancer context. The polyclonal format captures a spectrum of genetic modifications within the cell pool, offering a heterogeneous population suitable for pooled functional studies.
The parental NCI-H1975 cell line originates from a non-smoking female patient with non-small cell lung adenocarcinoma and harbors key oncogenic mutations, including EGFR L858R/T790M and PIK3CA G118D. These cells grow as adherent monolayers with epithelial morphology and display features of epithelial-mesenchymal transition, making them a clinically relevant model for EGFR-mutant lung adenocarcinoma and acquired resistance to tyrosine kinase inhibitors.
HECA functions as a transcriptional co-regulator and a structural component of the CCR4-NOT deadenylase complex, which includes CNOT1, CNOT2, CNOT3, CCR4, and CAF1. In the Notch signaling cascade, the NICD/CSL complex activates HECA expression, and HECA in turn modulates the stability of mRNAs encoding cell cycle regulators such as Cyclin D1 and CDKN1A (p21). Through interactions with this deadenylase machinery, HECA contributes to fine-tuning the G1/S transition, ultimately influencing E2F1-mediated transcription downstream of pRb. This positions HECA at the intersection of Notch-mediated transcriptional regulation and post-transcriptional control of proliferation-associated transcripts.
Disruption of HECA in the NCI-H1975 background is expected to perturb Notch-dependent transcriptional programs and destabilize cell cycle gene expression networks, potentially unmasking the tumor suppressor-like functions of HECA in this EGFR/PIK3CA-mutant adenocarcinoma model. The resulting dysregulation of the Cyclin D1/CDK4/CDK6?CpRb?CE2F1 axis may alter proliferative capacity, drug sensitivity, and invasive properties, providing a platform to explore mechanisms underlying resistance to EGFR-targeted therapies.
This product is suited for a range of applications, including investigation of Notch signaling in lung cancer, functional analysis of HECA in cell cycle control, and drug sensitivity profiling with EGFR tyrosine kinase inhibitors. Researchers can employ assays such as western blotting for HECA and downstream targets, RT-qPCR for Cyclin D1 and p21 expression, proliferation and cell cycle flow cytometry, RNA-seq for transcriptome-wide changes, and co-immunoprecipitation to assess CCR4-NOT complex integrity. Migration, invasion, and apoptosis assays further extend the utility of these knockout cells in exploring the multifaceted roles of HECA in tumor biology. For additional technical specifications, please contact Ascent Research.
