Blockade of CXCR4 in oral squamous cell carcinoma inhibits lymph node metastases
Daisuke Uchida1, Tomitaro Onoue, Nobuyuki Kuribayashi
1Department of Oral Surgery, Subdivision of Molecular Oral Medicine, Division of Integrated Sciences of Translational Research, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kuramoto, Tokushima, Japan. daisuke@dent.tokushima-u.ac.jp
Abstract:
We have previously demonstrated that a stromal cell-derived factor-1 (SDF-1; CXCL12)/CXCR4 system is involved in the establishment of lymph node metastasis in oral squamous cell carcinoma (OSCC). In this study, we investigated whether the blockade of CXCR4 inhibits lymph node metastasis in B88 OSCC cells. These cells harbour a functional CXCR4 and have the potential to metastasise to the lymph node in vivo. Following introduction of a vector that expresses short hairpin small interfering RNA (shRNA) against CXCR4, we isolated three clones (shCXCR4-16, -17 and -21) that showed decreased expression of CXCR4 mRNA. These clones also had reduced CXCR4 protein levels and showed impairments in calcium flux and cell migration in response to SDF-1. These cells were orthotopically inoculated into the masseter muscle of nude mice. Lymph node metastases, loss in body weight and tumour volumes were significantly inhibited in mice inoculated with shCXCR4-17 cells compared to mice inoculated with control cells. SDF-1-induced migration of B88 cells was significantly inhibited in vitro by the treatment with 1,1'-[1,4-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane octahydrochloride (AMD3100), a CXCR4 antagonist. Subcutaneous administration of AMD3100 significantly inhibited the lymph node metastases of B88 cells when they were orthotopically inoculated into the masseter muscle of nude mice. Moreover, the enhanced production of interleukin (IL)-6 and IL-8 in response to SDF-1 was inhibited by shRNA against CXCR4 or by treatment with AMD3100. These results suggest that blockade of CXCR4 may be a potent anti-metastatic therapy against lymph node metastases in cases of CXCR4-related OSCC.
Insights
Blocking the CXCR4 receptor effectively inhibits lymph node metastasis in oral squamous cell carcinoma (OSCC). This research shows CXCR4 blockade as a promising anti-metastatic therapy for OSCC patients.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Metastasis Research
Background:
- The stromal cell-derived factor-1 (SDF-1)/CXCR4 system plays a role in oral squamous cell carcinoma (OSCC) lymph node metastasis.
- B88 OSCC cells possess functional CXCR4 and the capacity for lymph node metastasis in vivo.
Purpose of the Study:
- To investigate the efficacy of blocking CXCR4 in inhibiting lymph node metastasis of B88 OSCC cells.
- To evaluate the anti-metastatic potential of CXCR4 blockade in an in vivo model of OSCC.
Main Methods:
- Generated OSCC cell clones with reduced CXCR4 expression using short hairpin small interfering RNA (shRNA).
- Assessed CXCR4 expression, calcium flux, and cell migration in response to SDF-1.
- Orthotopically inoculated modified OSCC cells into nude mice to evaluate lymph node metastasis.
- Administered AMD3100, a CXCR4 antagonist, to assess its effect on metastasis and cytokine production.
Main Results:
- OSCC clones with reduced CXCR4 expression exhibited impaired calcium flux and migration.
- Lymph node metastases, body weight loss, and tumor volumes were significantly inhibited in mice treated with shCXCR4 cells.
- AMD3100 treatment significantly inhibited SDF-1-induced cell migration in vitro and lymph node metastases in vivo.
- Blockade of CXCR4 or AMD3100 treatment reduced SDF-1-induced interleukin (IL)-6 and IL-8 production.
Conclusions:
- CXCR4 blockade, achieved through shRNA or AMD3100, effectively inhibits lymph node metastasis in OSCC.
- Targeting the CXCR4 pathway presents a potential anti-metastatic therapeutic strategy for CXCR4-related OSCC.
