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Updated: Dec 14, 2025
Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment
Published on: April 22, 2019
An HNSCC syngeneic mouse model for tumor immunology research and preclinical evaluation
You Fu1, Guocai Tian1, Jiang Li1
1Shanghai Key Laboratory of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China.
Abstract:
The lack of reliable animal models to assess the safety and efficacy of drugs and to explore the underlying molecular mechanisms is one of the most severe impediments in head and neck squamous cell carcinoma (HNSCC) tumor immunology research. The majority of xenograft tumor models established using immunodeficient mice neglect the effects of T cells. To date, to the best of our knowledge, there is no syngeneic tumor model available that reflects the immune microenvironmental features of HNSCC tumors. To solve this issue, the present study used 4‑nitroquinoline‑1‑oxide (4‑NQO) to induce squamous cell carcinoma in C57BL/6 mice. Three HNSCC cell lines were then established, and one of these, termed JC1, was selected for further analysis due to its enhanced proliferative ability and tumorigenicity in immunodeficient nude mice. However, none of the 3 cell lines could form tumors in immunocompetent mice. Due to the different tumorigenicities in nude and C57BL/6 mice, the immune system may play an important role in inoculated JC1 tumor progression. Chemical induction was used to establish the tumorigenicity‑enhanced cell line, JC1‑2, which can form syngeneic tumors in immunocompetent C57BL/6 mice. Next‑generation sequencing (NGS) was used to perform the immunogenomic and transcriptomic characterization of the JC1‑2 cells. Splenocytes were isolated from C57BL/6 mice and co‑cultured with JC1‑2 cells to verify the responsiveness of the interferon (IFN)‑γ pathway in the JC1‑2 cell line. Unlike the majority of syngeneic mouse tumors, the JC1‑2‑formed tumors resembled 'inflamed tumors' due to the abundancy of immune cells in the tumor microenvironment. Moreover, more intense immune responses were observed in the orthotopic mouse model than in the heterotopic model. Thus, this model could be used to delineate the interactions between HNSCC and lymphocytes, and to validate novel immunotherapy targets.
Insights
Researchers developed a new syngeneic mouse model for head and neck squamous cell carcinoma (HNSCC) research. This model, using JC1-2 cells, accurately reflects the tumor microenvironment and immune responses, aiding in immunotherapy development.
Area of Science:
- Oncology
- Immunology
- Animal Models
Background:
- Head and neck squamous cell carcinoma (HNSCC) research is hindered by the lack of reliable animal models that incorporate immune system interactions.
- Existing xenograft models in immunodeficient mice do not account for T cell activity, a crucial component of tumor immunology.
- There is a significant need for a syngeneic HNSCC tumor model that mirrors the complex immune microenvironment of human tumors.
Purpose of the Study:
- To develop a novel, immunocompetent syngeneic animal model for head and neck squamous cell carcinoma (HNSCC) research.
- To establish and characterize a new HNSCC cell line (JC1-2) capable of forming tumors in immunocompetent mice.
- To utilize this model for studying HNSCC-immune cell interactions and identifying potential immunotherapy targets.
Main Methods:
- Induction of squamous cell carcinoma in C57BL/6 mice using 4-nitroquinoline-1-oxide (4-NQO).
- Establishment and selection of HNSCC cell lines, followed by chemical induction to enhance tumorigenicity in immunocompetent mice (JC1-2).
- Next-generation sequencing (NGS) for immunogenomic and transcriptomic profiling of JC1-2 cells; co-culture assays with splenocytes to assess interferon-gamma pathway responsiveness.
Main Results:
- A tumorigenic HNSCC cell line (JC1-2) was successfully established, capable of forming syngeneic tumors in immunocompetent C57BL/6 mice.
- JC1-2 tumors exhibited an 'inflamed' phenotype with abundant immune cells in the tumor microenvironment, unlike typical syngeneic tumors.
- Orthotopic models demonstrated more intense immune responses compared to heterotopic models, validating the model's utility in reflecting in vivo immune dynamics.
Conclusions:
- The developed JC1-2 syngeneic model provides a valuable tool for studying head and neck squamous cell carcinoma (HNSCC) tumor immunology.
- This model effectively recapitulates key features of the HNSCC immune microenvironment, including immune cell infiltration and inflammatory responses.
- The JC1-2 model is suitable for investigating HNSCC-lymphocyte interactions and for pre-clinical validation of novel immunotherapy strategies.
