NCI-H1568 Xenograft Model Service for NSCLC

The NCI-H1568 xenograft model, harboring the KRAS G12V oncogenic driver together with ATM and TP53 alterations, offers a physiologically relevant platform for interrogating KRAS-dependent tumor biology and therapeutic resistance in lung adenocarcinoma. Alfa Cytology delivers authenticated NCI-H1568 cell line-derived xenograft services with documented mutational fidelity, enabling robust preclinical evaluation of KRAS-targeted agents, downstream pathway inhibitors, and combination regimens in NSCLC.

Overview of NCI-H1568 Xenograft Model for NSCLC

NCI-H1568 was established in December 1986 from a lymph node metastasis of lung adenocarcinoma in a 48-year-old female patient of European descent. This epithelial cell line carries the KRAS G12V activating mutation while remaining wild-type for EGFR and ALK, a molecular profile representative of a substantial subset of treatment-naïve lung adenocarcinomas that exhibit primary resistance to EGFR-directed therapies. Additional alterations in ATM and TP53 further complicate its DNA damage response landscape, rendering the model particularly instructive for studying synthetic lethal interactions and combination strategies that exploit genomic instability.

Fig 1. In the NCI-H1568 (SMARCA4-deficient) xenograft model, a single subcutaneous dose of compound 6 reduced intratumoral SMARCA2 by 90% at 6 h (−78% at 48 h), translating into 77% and 84% tumor growth inhibition across two intermittent dosing regimens. (Kofink, Christiane, et al., 2022)

A distinctive feature of NCI-H1568 is its pronounced cellular heterogeneity, encompassing hierarchically organized subpopulations that range from stem-like holoclones to differentiated epithelial progeny. When propagated as a subcutaneous xenograft, this line recapitulates the clonal diversity observed in vitro, forming tumors that retain KRAS-driven signaling alongside variable expression of stemness and lineage-specific markers. Consequently, the NCI-H1568 xenograft serves as an excellent system for assessing compounds that target KRAS-mutant tumor maintenance, disrupt cancer stem cell reservoirs, or modulate the tumor microenvironment in NSCLC.

Cell Line Information: NCI-H1568

The NCI-H1568 cell line was deposited by Dr. A. Gazdar and Dr. J. Minna and is available through ATCC. Its well-documented genotype and phenotypic traits are summarized below:

Our Services

Alfa Cytology maintains NCI-H1568 under authenticated culture conditions and constructs xenografts with verified KRAS G12V status, offering study designs that encompass monotherapy efficacy, combination arm screening, and longitudinal stemness marker profiling to advance your KRAS-directed NSCLC pipeline.

Workflow of NCI-H1568 Xenograft Model Construction

NCI-H1568 xenograft construction follows a protocol tailored to its adherent epithelial phenotype and heterogeneous clonal architecture, ensuring consistent engraftment and interpretable pharmacological readouts. The workflow proceeds through six sequential stages:

1. Cell Culture and Batch Authentication: NCI-H1568 cells are expanded in RPMI-1640 with 10% FBS. Prior to implantation, each batch undergoes STR profiling, mycoplasma testing, and KRAS G12V mutational confirmation to guarantee genetic integrity.
2. Harvest and Matrigel Suspension: Confluent monolayers are detached with Accutase, washed, and counted. Cells are resuspended at 1 × 10⁷ cells/mL in a 1:1 blend of serum-free RPMI-1640 and high-concentration Matrigel, preserving the clonal heterogeneity essential for tumor engraftment.
3. Subcutaneous Implantation: A 100 µL inoculum containing 1 × 10⁶ viable cells is injected subcutaneously into the flank of female athymic nude mice. Tumor nodules generally become palpable within 12–16 days post-injection.
4. Tumor Monitoring and Group Allocation: Tumor dimensions are measured twice weekly with digital calipers, and volume is calculated as (length × width²) / 2. Once tumors reach 100–150 mm³, mice are randomized into treatment cohorts matched for tumor volume and body weight.
5. Compound Administration and In-Life Monitoring: Test articles are dosed according to the predefined schedule. Tumor volumes and body weights are recorded at regular intervals. Optional blood sampling permits pharmacokinetic and pharmacodynamic correlation.
6. Endpoint Analysis and Tissue Processing: At study termination, tumors are excised, weighed, and photographed. Tissues are fixed in 10% neutral buffered formalin for H&E and immunohistochemistry, with aliquots snap-frozen for genomic, transcriptomic, or proteomic analyses.

Fig 2. NCI-H1568 xenograft model construction workflow.

Case Study-NCI-H1568 Xenograft Model Development

In a recent preclinical investigation, the NCI-H1568 xenograft was utilized to evaluate the antitumor potential of a novel KRAS G12V-selective small molecule inhibitor administered in combination with a downstream MEK antagonist. Following subcutaneous engraftment in immunodeficient mice, animals with established tumors were randomized and treated for 28 days. The combination regimen produced marked tumor growth inhibition relative to vehicle and single-agent controls, with responder tumors showing reduced phospho-ERK staining and diminished Ki-67 proliferation indices upon post-study immunohistochemical analysis. These preclinical observations provided rationale for extended tolerability and biomarker-driven patient selection strategies in subsequent development stages.

Why Choose Alfa Cytology?

Alfa Cytology integrates molecular authentication with flexible study execution to deliver NCI-H1568 xenograft data that advance your KRAS-mutant NSCLC therapeutic program.

• Every NCI-H1568 batch is authenticated by STR analysis and confirmed for KRAS G12V mutational status prior to study initiation.
• We offer customizable combination study designs that pair KRAS inhibitors with MEK, ERK, or immunomodulatory agents.
• Tumor growth is monitored with electronic calipers and optional non-invasive imaging for continuous longitudinal assessment.
• All procedures are conducted in AAALAC-accredited facilities under IACUC-approved protocols with dedicated veterinary care.
• Study deliverables encompass statistical reports, individual animal tracking, and publication-ready histopathology documentation.

Contact Us

If your NSCLC drug discovery program requires a validated KRAS-mutant preclinical model with documented clonal heterogeneity, reach out to Alfa Cytology today. Our preclinical specialists will work with you to configure an NCI-H1568 xenograft study that aligns with your target indication, compound mechanism, and regulatory submission strategy.

For research use only. Not intended for any clinical use.