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JunB promotes cell invasion, migration and distant metastasis of head and neck squamous cell carcinoma
Hiroshi Hyakusoku1, Daisuke Sano2,3, Hideaki Takahashi4,5
1Department of Biology and Function in Head and Neck, Yokohama City University Graduate School of Medicine, Yokohama, Japan. hhyaku@yokohama-cu.ac.jp.
Background:
While treatment failure in cases of head and neck squamous cell carcinoma (HNSCC) frequently takes the form of locoregional recurrences and distant metastasis, our understanding of the mechanisms of metastasis in HNSCC is limited. We initially performed the upstream and key nodes analysis together with whole gene microarray analysis characterized by distant metastatic potential in vivo with HNSCC cell lines and identified JunB, a member of the activator protein-1 (AP-1) family, as a key molecule in the regulation of the pathways related to distant metastasis in HNSCC. We have therefore tested the hypothesis that JunB plays a crucial role in distant metastasis in HNSCC.
Methods:
To study the role of JunB on metastatic potential of HNSCC, small interfering RNA (siRNA)-mediated knockdown and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (cas9) system (CRISPR/Cas9)-mediated knockout of JunB in HNSCC cells were established and the abilities of cell invasion and migration in vitro were examined. The efficacy of knockout of JunB was also examined using an experimental lung metastatic mouse model of HNSCC. In addition, to study if the role of JunB in HNSCC cell migration and invasiveness is related to epithelial-to-mesenchymal transition (EMT), cell morphology and expression of mesenchymal or epithelial marker on siRNA mediated JunB knockdown in HNSCC cells were examined with or without TGF-β stimulation.
Results:
siRNA knockdown and sgRNA knockout of JunB in metastatic HNSCC cells significantly suppressed both cell invasion and migration in vitro. In addition, the knockout of JunB in metastatic HNSCC cells significantly repressed the incidence of lung metastases and prolonged the survival in vivo. However, we did not observe any change in cell morphology with the down-regulation of mesenchymal markers and up-regulation of epithelial markers in response to siRNA-mediated JunB knockdown in HNSCC cells.
Conclusion:
These results suggested that JunB could play an important role in promoting cell invasion, migration and distant metastasis in HNSCC via pathways other than EMT and that the down-regulation of JunB may become an effective strategy for patients with invasive HNSCC.
Insights
JunB promotes head and neck squamous cell carcinoma (HNSCC) metastasis through pathways independent of epithelial-mesenchymal transition (EMT). Down-regulating JunB may offer a new strategy against invasive HNSCC.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Metastasis Research
Background:
- Head and neck squamous cell carcinoma (HNSCC) often recurs or metastasizes, but metastasis mechanisms remain unclear.
- JunB, an activator protein-1 (AP-1) family member, was identified as a key regulator of HNSCC distant metastasis.
- This study investigates the critical role of JunB in promoting HNSCC distant metastasis.
Purpose of the Study:
- To elucidate the function of JunB in HNSCC cell invasion and migration.
- To determine JunB's role in HNSCC metastasis in vivo.
- To explore the relationship between JunB and epithelial-to-mesenchymal transition (EMT) in HNSCC.
Main Methods:
- JunB was knocked down using small interfering RNA (siRNA) and knocked out using CRISPR/Cas9 in HNSCC cells.
- In vitro cell invasion and migration assays were performed.
- An experimental lung metastasis mouse model was used to assess JunB's in vivo efficacy.
- Cell morphology and marker expression were analyzed to evaluate EMT involvement.
Main Results:
- JunB knockdown and knockout significantly reduced HNSCC cell invasion and migration in vitro.
- JunB knockout suppressed lung metastasis incidence and improved survival in vivo.
- No significant changes in cell morphology or EMT markers were observed upon JunB down-regulation.
Conclusions:
- JunB promotes HNSCC cell invasion, migration, and distant metastasis through mechanisms independent of EMT.
- Targeting JunB could be a promising therapeutic strategy for invasive HNSCC.
- Further research into JunB-mediated pathways is warranted for HNSCC treatment development.
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