NCI-H2009 Xenograft Model Service for NSCLC

The NCI-H2009 xenograft model, carrying the KRAS G12A oncogenic driver together with mutant TP53 and high TTF-1 expression, provides a robust preclinical platform for evaluating targeted therapies and combination regimens in lung adenocarcinoma. Alfa Cytology delivers authenticated NCI-H2009 cell line-derived xenograft services with documented mutational fidelity and TRU-type marker profiling, enabling precise assessment of KRAS-directed agents and downstream pathway inhibitors in NSCLC.

Overview of NCI-H2009 Xenograft Model for NSCLC

NCI-H2009 was established in August 1988 from a metastatic lymph node of a 68-year-old Caucasian female with stage IV lung adenocarcinoma. The patient had a 30 pack-year smoking history and had received prior chemotherapy and radiation therapy. This epithelial cell line harbors the KRAS G12A activating mutation in a wild-type EGFR background, a molecular profile representative of the approximately 7.6% of lung adenocarcinomas driven by this specific KRAS variant. Additional TP53 mutation and high TTF-1 (NKX2-1) expression further define its identity as a TRU-type (terminal respiratory unit) lung adenocarcinoma model with characteristic sensitivity to aurora kinase inhibition and MEK blockade.

Fig 1. CRIF1 promoted SIRT3-mediated deacetylation of PYCR1 at K228 in NCI-H2009 cells, and CRIF1 knockdown suppressed PYCR1 activity — the mechanistic basis for CRIF1-driven growth of NCI-H2009 xenograft tumors. (Wang, Qi, et al., 2022)

When propagated as a subcutaneous xenograft in immunodeficient mice, NCI-H2009 forms moderately aggressive tumors with a doubling time of approximately 29.8 hours in vitro and consistent engraftment in vivo. The model exhibits pronounced chromosomal instability with extensive structural rearrangements and double-minute chromosomes, recapitulating the genomic complexity of smoking-associated lung adenocarcinoma. Its well-documented resistance to cisplatin and sensitivity to combined FGFR1/PLK1 inhibition make it an invaluable system for benchmarking novel KRAS-mutant therapeutic strategies and studying acquired resistance mechanisms.

Cell Line Information: NCI-H2009

The NCI-H2009 cell line was deposited by Dr. A. Gazdar and Dr. J. Minna and is distributed by ATCC. Its extensively characterized genotype and phenotype support high-confidence preclinical translation:

Our Services

Alfa Cytology maintains NCI-H2009 under authenticated culture conditions and constructs xenografts with verified KRAS G12A and TP53 mutational status, offering study designs that encompass monotherapy efficacy, combination arm screening, and longitudinal TTF-1 expression profiling to advance your KRAS-mutant NSCLC pipeline.

Workflow of NCI-H2009 Xenograft Model Construction

NCI-H2009 xenograft construction follows a standardized protocol optimized for adherent epithelial cells with KRAS-driven signaling, ensuring reproducible engraftment and robust pharmacological readouts. The workflow proceeds through six integrated phases:

1. Cell Expansion and Authentication: NCI-H2009 cells are propagated in RPMI-1640 with 10% FBS to log-phase growth. Each batch is verified by STR profiling, mycoplasma PCR, and KRAS G12A mutational analysis prior to implantation.
2. Harvest and Matrigel Preparation: Adherent cultures are detached with trypsin-EDTA, washed, and counted. Cells are resuspended at 1 × 10⁷ cells/mL in a 1:1 mixture of serum-free RPMI-1640 and high-concentration Matrigel to support initial tumor stroma formation.
3. Subcutaneous Engraftment: A 100 µL suspension containing 1 × 10⁶ viable cells is injected subcutaneously into the flank of female athymic nude mice. Tumor nodules typically become palpable within 7–10 days post-injection.
4. Tumor Monitoring and Cohort Randomization: Tumor dimensions are measured twice weekly with electronic calipers, with volume derived from the formula (length × width²) / 2. Upon reaching 100–150 mm³, mice are randomized into treatment groups stratified by tumor volume and body weight.
5. Dosing and In-Life Assessments: Test compounds are administered via the designated route and schedule. Tumor volumes and body weights are recorded at defined intervals throughout the dosing phase. Optional blood draws support PK/PD correlation analysis.
6. Terminal Procedures and Sample Processing: At study endpoint, tumors are excised, weighed, and imaged. Tissues are fixed in 10% neutral buffered formalin for H&E and IHC analysis, with portions snap-frozen in liquid nitrogen for downstream genomic or proteomic studies.

Fig 2. NCI-H2009 xenograft model construction workflow.

Case Study-NCI-H2009 Xenograft Model Development

In a preclinical program evaluating a novel MEK1/2 inhibitor in combination with a repurposed FLT3 kinase antagonist, NCI-H2009 xenografts were established in athymic nude mice and dosed orally once daily for 21 days. The combination regimen produced dose-dependent tumor growth inhibition across multiple treatment arms, with the highest dose cohort demonstrating sustained suppression relative to vehicle-treated controls. Post-study immunohistochemistry revealed diminished phospho-ERK staining and elevated cleaved caspase-3 expression in responder tumors, alongside preserved TTF-1 positivity confirming TRU-type lineage maintenance. These preclinical observations provided mechanistic validation and supported the compound's advancement into extended tolerability and biomarker-driven patient selection strategies.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for NCI-H2009 xenograft studies ensures access to a genetically characterized TRU-type model managed under rigorous quality standards.

• Every NCI-H2009 batch is validated by STR analysis and confirmed for KRAS G12A and TP53 mutational status before study initiation.
• We provide flexible dosing paradigms including oral gavage, intravenous, and intraperitoneal administration with customizable frequency and duration.
• Tumor growth data are captured with electronic calipers and optional bioluminescence imaging for real-time longitudinal monitoring.
• All in vivo work is performed in AAALAC-accredited vivaria under IACUC-reviewed protocols with full veterinary support.
• Comprehensive study reports include statistical analysis, individual animal data, and publication-quality histopathology images with TTF-1 staining.

Contact Us

To advance your KRAS-mutant lung adenocarcinoma therapeutic candidate through a validated TRU-type preclinical model, contact Alfa Cytology today. Our scientific team will collaborate with you to design a tailored NCI-H2009 xenograft program that aligns with your target product profile and regulatory milestones.

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