O-charoenrat Pornchai, Leksrisakul Piyavadee, Sangruchi Supatra
Department of Head and Neck Surgery, Siriraj Hospital Medical School, Mahidol University, Bangkok, Thailand.
Int J Cancer. 2006 May 15;118(10):2548-53. doi: 10.1002/ijc.21644.
Matrix metalloproteinases (MMPs) play an important role in several steps of cancer development. A single guanine insertion polymorphism (2G) in the MMP1 promoter sequence at -1,607 creates an Ets binding site and thus results in enhancing transcriptional activity. This study aimed to evaluate the contribution of this 2G polymorphism on susceptibility and aggressiveness of HNSCC. A panel of HNSCC cell lines and peritumoral fibroblasts were examined for the MMP1 genotypes and expression levels. Genomic DNA was extracted from peripheral blood of 300 patients with newly diagnosed HNSCC and from 300 age- and gender-matched cancer-free controls. Genotyping was carried out using a PCR-RFLP assay. The levels of MMP1 mRNA expression were evaluated by the quantitative RT-PCR and a correlation with different genotype was determined. Odds ratio (OR) for cancer risk were calculated using multivariate logistic regression. In addition, a correlation between the 2G/2G genotype and clinicopathological parameters was examined. Eleven out of 18 HNSCC cell lines showed the 2G/2G genotype (61%) and only 1 cell line had the 1G/1G genotype (5.6%). Cell lines with the 2G/2G genotype expressed significantly higher mean MMP1 mRNA level than those with other genotypes. In clinical model, subjects carrying the homozygous 2G/2G genotype had a higher risk of head and neck cancer compared with subjects with other genotypes (adjusted OR: 2.28; 95% CI: 1.58-3.27), controlling for major confounders. A correlation between promoter polymorphisms and the levels of MMP1 expression in cancer tissues was found, and this 2G/2G genotype was correlated with the adverse clinicopathological parameters. Finally, the highest level of MMP1 enhancement was demonstrated in the coculture of tumor cells and peritumoral fibroblasts of 2G homozygotes. These findings suggest that the presence of 2G polymorphism at the MMP1 promoter is associated with the development and progression of HNSCC.
基质金属蛋白酶（MMPs）在癌症发展的多个步骤中发挥着重要作用。MMP1启动子序列-1607处的单个鸟嘌呤插入多态性（2G）产生了一个Ets结合位点，从而导致转录活性增强。本研究旨在评估这种2G多态性对HNSCC易感性和侵袭性的影响。对一组HNSCC细胞系和肿瘤周围成纤维细胞进行了MMP1基因型和表达水平检测。从300例新诊断的HNSCC患者的外周血以及300例年龄和性别匹配的无癌对照者的外周血中提取基因组DNA。采用PCR-RFLP分析法进行基因分型。通过定量RT-PCR评估MMP1 mRNA表达水平，并确定其与不同基因型的相关性。使用多因素逻辑回归计算癌症风险的优势比（OR）。此外，还研究了2G/2G基因型与临床病理参数之间的相关性。18个HNSCC细胞系中有11个显示2G/2G基因型（61%），只有1个细胞系具有1G/1G基因型（5.6%）。具有2G/2G基因型的细胞系平均MMP1 mRNA水平明显高于其他基因型的细胞系。在临床模型中，与其他基因型的受试者相比，携带纯合2G/2G基因型的受试者患头颈癌的风险更高（校正OR：2.28；95%CI：1.58 - 3.27），同时控制了主要混杂因素。发现启动子多态性与癌组织中MMP1表达水平之间存在相关性，并且这种2G/2G基因型与不良临床病理参数相关。最后，在2G纯合子的肿瘤细胞与肿瘤周围成纤维细胞的共培养中，MMP1增强水平最高。这些发现表明，MMP1启动子处2G多态性的存在与HNSCC的发生和发展有关。
