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Updated: Jun 20, 2025
In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System
Published on: January 9, 2020
Human papillomavirus-associated head and neck squamous cell carcinoma cells lose viability during triggered myocyte
Sarah Gendreizig1, Laura Martínez-Ruiz2,3,4, Alba López-Rodríguez2,3,4
1Department of Otolaryngology, Head and Neck Surgery, Campus Klinikum Bielefeld Mitte, University Hospital OWL of Bielefeld University, Teutoburger Str. 50, 33604, Bielefeld, Germany.
Abstract:
Head and neck squamous cell carcinoma (HNSCC) is a highly malignant disease, and death rates have remained at approximately 50% for decades. New tumor-targeting strategies are desperately needed, and a previous report indicated the triggered differentiation of HPV-negative HNSCC cells to confer therapeutic benefits. Using patient-derived tumor cells, we created a similar HNSCC differentiation model of HPV+ tumor cells from two patients. We observed a loss of malignant characteristics in differentiating cell culture conditions, including irregularly enlarged cell morphology, cell cycle arrest with downregulation of Ki67, and reduced cell viability. RNA-Seq showed myocyte-like differentiation with upregulation of markers of myofibril assembly. Immunofluorescence staining of differentiated and undifferentiated primary HPV+ HNSCC cells confirmed an upregulation of these markers and the formation of parallel actin fibers reminiscent of myoblast-lineage cells. Moreover, immunofluorescence of HPV+ tumor tissue revealed areas of cells co-expressing the identified markers of myofibril assembly, HPV surrogate marker p16, and stress-associated basal keratinocyte marker KRT17, indicating that the observed myocyte-like in vitro differentiation occurs in human tissue. We are the first to report that carcinoma cells can undergo a triggered myocyte-like differentiation, and our study suggests that the targeted differentiation of HPV+ HNSCCs might be therapeutically valuable.
Insights
Researchers triggered a myocyte-like differentiation in human papillomavirus-positive head and neck squamous cell carcinoma (HPV+ HNSCC) cells. This differentiation reduced malignant traits, suggesting a potential new therapeutic strategy for HNSCC.
Area of Science:
- Oncology
- Cell Biology
- Cancer Research
Background:
- Head and neck squamous cell carcinoma (HNSCC) has a high mortality rate.
- Current treatments lack efficacy, necessitating novel therapeutic strategies.
- Previous studies suggested therapeutic benefits from differentiating human papillomavirus-negative (HPV-negative) HNSCC cells.
Purpose of the Study:
- To investigate the potential for triggered myocyte-like differentiation in HPV-positive (HPV+) HNSCC.
- To determine if this differentiation impacts malignant characteristics of HPV+ HNSCC.
- To explore the therapeutic implications of inducing differentiation in HPV+ HNSCC.
Main Methods:
- Established a differentiation model using patient-derived HPV+ HNSCC cells.
- Analyzed changes in cell morphology, cell cycle (Ki67), and viability.
- Utilized RNA-sequencing (RNA-Seq) to identify differentiation markers.
- Performed immunofluorescence staining on cell cultures and tumor tissues to confirm marker expression and cellular changes.
Main Results:
- Differentiating HPV+ HNSCC cells exhibited reduced malignancy, including altered morphology, cell cycle arrest, and decreased viability.
- RNA-Seq revealed upregulation of myocyte-like differentiation markers, including those for myofibril assembly.
- Immunofluorescence confirmed increased expression of these markers and formation of actin fibers, resembling myoblasts.
- In HPV+ tumor tissue, cells co-expressed differentiation markers, p16 (HPV marker), and KRT17 (keratinocyte marker), indicating in vivo differentiation.
Conclusions:
- This study is the first to report triggered myocyte-like differentiation in carcinoma cells.
- The observed differentiation in HPV+ HNSCC cells reduced their malignant phenotype.
- Targeted induction of this myocyte-like differentiation may represent a promising therapeutic approach for HPV+ HNSCC.
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