趋化因子受体CXCR4及其配体CXCL12在口腔鳞状细胞癌增殖和迁移过程中的作用

[The role of chemokine receptor CXCR4 and its ligand CXCL12 in the process of proliferation and migration of oral squamous cell carcinoma].

作者信息

Yin Dong, Zhang Zuo, Gao Su, Li Bin

机构信息

Dept. of Stomatology, The People's Hospital of Ningxia Hui Autonomous Region, Yinchuan 750021, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2013 Feb;31(1):8-12.

PMID:23484292

Abstract

OBJECTIVE

To investigate the expression of chemokine receptor CXCR4 and its ligand CXCL12 in oral squamous cell carcinoma (OSCC) cells, and their effects on proliferation and migration of tumor cells.

METHODS

The expression of CXCR4 mRNA and protein in 20 cases of OSCC tissue, cervical lymph nodes, tongue cancer cell line Tca8113, buccal cancer cell line Bca885, and 10 specimens of normal oral mucosa tissue were examined by reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blot. The expression of CXCL12 mRNA was examined by RT-PCR. MTT assay was used to evaluate the CXCR4/CXCL12 influence on proliferation of tumor cells. Chemotaxis and migration response to CXCR4 particular ligand-CXCL12 were detected by chemotaxis assay.

RESULTS

Expression levels of CXCR4 mRNA and protein in OSCC tissues, Tca8113 and Bca885 cells were 2.31 +/- 1.13, 1.89 +/- 1.20, 1.67 +/- 1.10 and 1.36 +/- 0.15, 1.85 +/- 0.34, 1.97 +/- 0.23, respectively. CXCR4 mRNA and protein couldn't be detected in normal tissues. CXCL12 mRNA level in cervical lymph nodes was 1.14 +/- 0.87, CXCL12 mRNA couldn't be detected in OSCC and normal tissues. 2) In MTT assay, recombinant CXCL12 stimulated proliferation of tumor cells and CXCR4 neutralization by monoclonal antibodies decreased proliferation. 3) The chemotactic migration of OSCC cells could be induced by CXCL12. CXCL12 at a concentration between 30 ng x mL(-1) and 100 ng x mL(-1) induced the chemotactic migration of OSCC cells in a dose-dependent manner.

CONCLUSION

CXCR4/CXCL12 system may promote proliferation and migration of tumor cells, and may play an important role in lymph node metastasis of OSCC.

摘要

目的

研究趋化因子受体CXCR4及其配体CXCL12在口腔鳞状细胞癌（OSCC）细胞中的表达情况，以及它们对肿瘤细胞增殖和迁移的影响。

方法

采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测20例OSCC组织、颈部淋巴结、舌癌细胞系Tca8113、颊癌细胞系Bca885及10例正常口腔黏膜组织中CXCR4 mRNA和蛋白的表达。采用RT-PCR检测CXCL12 mRNA的表达。采用MTT法评估CXCR4/CXCL12对肿瘤细胞增殖的影响。采用趋化实验检测OSCC细胞对CXCR4特异性配体CXCL12的趋化和迁移反应。

结果

1）OSCC组织、Tca8113和Bca885细胞中CXCR4 mRNA和蛋白的表达水平分别为2.31±1.13、1.89±1.20、1.67±1.10和1.36±0.15、1.85±0.34、1.97±0.23。正常组织中未检测到CXCR4 mRNA和蛋白。颈部淋巴结中CXCL12 mRNA水平为1.14±0.87，OSCC组织和正常组织中未检测到CXCL12 mRNA。2）在MTT实验中，重组CXCL12可刺激肿瘤细胞增殖，单克隆抗体中和CXCR4可降低增殖。3）CXCL12可诱导OSCC细胞的趋化迁移。浓度在30 ng·mL-1至100 ng·mL-1之间的CXCL12可诱导OSCC细胞的趋化迁移呈剂量依赖性。