Department 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.
Eur J Cancer. 2011 Feb;47(3):452-9. doi: 10.1016/j.ejca.2010.09.028. Epub 2010 Oct 19.
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.
我们之前已经证明，基质细胞衍生因子-1（SDF-1；CXCL12）/CXCR4 系统参与了口腔鳞状细胞癌（OSCC）淋巴结转移的建立。在这项研究中，我们研究了阻断 CXCR4 是否可以抑制 B88 OSCC 细胞的淋巴结转移。这些细胞具有功能性的 CXCR4，并且具有体内转移到淋巴结的潜力。在引入表达针对 CXCR4 的短发夹 RNA（shRNA）的载体后，我们分离出三个显示 CXCR4 mRNA 表达降低的克隆（shCXCR4-16、-17 和 -21）。这些克隆还表现出 CXCR4 蛋白水平降低，并在对 SDF-1 的钙通量和细胞迁移方面表现出缺陷。这些细胞被原位接种到裸鼠的咬肌中。与接种对照细胞的小鼠相比，接种 shCXCR4-17 细胞的小鼠的淋巴结转移、体重减轻和肿瘤体积明显受到抑制。SDF-1 诱导的 B88 细胞迁移在体外通过用 CXCR4 拮抗剂 1,1'-[1,4-亚苯基双（亚甲基）]双-1,4,8,11-四氮杂环十四烷八盐酸盐（AMD3100）处理而显著抑制。AMD3100 皮下给药可显著抑制当 B88 细胞原位接种到裸鼠的咬肌时的淋巴结转移。此外，shRNA 针对 CXCR4 或用 AMD3100 处理可抑制对 SDF-1 的反应中白细胞介素（IL）-6 和 IL-8 的增强产生。这些结果表明，阻断 CXCR4 可能是针对 CXCR4 相关 OSCC 淋巴结转移的有效抗转移治疗方法。
