The HTATIP2 Knockout NCI-H1975 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human lung adenocarcinoma NCI-H1975 cell line, engineered to disrupt the HTATIP2 gene (HIV-1 Tat interactive protein 2, also known as TIP30). This loss-of-function model enables systematic investigation of HTATIP2-mediated tumor suppression in a genetically defined non-small cell lung cancer (NSCLC) context without selecting for single-cell clones, preserving polyclonal heterogeneity that better reflects biological variability.
The NCI-H1975 host cell line is a well-established model of NSCLC, carrying both an activating EGFR L858R mutation and the secondary T790M gatekeeper mutation. The T790M substitution confers resistance to first- and second-generation EGFR tyrosine kinase inhibitors (TKIs), making this line particularly valuable for dissecting mechanisms of acquired drug resistance and for evaluating next-generation EGFR-targeted therapeutics in clinically relevant settings.
HTATIP2 is a multifunctional tumor suppressor that operates at the crossroads of several oncogenic signaling axes. It directly binds and inhibits Src kinase activity and associates with importin beta to block nuclear transport of beta-catenin, thereby repressing Wnt target genes such as cyclin D1 and c-Myc. This action curbs cell cycle progression, migration, and angiogenesis while sensitizing cells to caspase-3-mediated apoptosis. HTATIP2 also interacts with nucleoporins, estrogen receptor alpha, and Axin, and its expression is governed by upstream inputs including STAT3, c-Myc, estrogen, microRNA-182, and promoter methylation, coupling transcriptional, epigenetic, and hormonal regulation to nucleocytoplasmic shuttling.
In the NCI-H1975 background, disruption of HTATIP2 is expected to relieve inhibition of Src/FAK and Wnt/beta-catenin pathways, which are frequently hyperactivated in EGFR TKI-resistant NSCLC and contribute to enhanced invasive and metastatic phenotypes. Thus, this polyclonal knockout population offers a physiologically relevant platform to examine how loss of a key tumor suppressor synergizes with oncogenic EGFR signaling to drive lung adenocarcinoma progression, providing critical insights into potential combinatorial therapeutic vulnerabilities.
These HTATIP2 knockout cells facilitate a wide array of functional assays, including MTT/CCK-8 proliferation assays, Transwell migration and invasion studies, Annexin V/PI apoptosis analyses, and Western blotting for downstream effectors like cyclin D1, c-Myc, and phosphorylated Src. Researchers can employ Src kinase activity assays, immunofluorescence to visualize beta-catenin subcellular localization, and colony formation assays to assess clonogenic potential. The model is also suitable for in vivo xenograft experiments to probe tumor suppression and drug response in an EGFR-mutant milieu. For further technical details or custom requests, please contact Ascent Research.
