MMP-13與TIMP-1基因多型性與台灣口腔癌前病變癌化潛力和鱗狀細胞癌發生之相關性探討

Susceptibility to Oral Squamous Cell Carcinoma and Malignance Potential and Genetic Polymorphisms of MMP-13 and TIMP-1 Genes in a Taiwan Population

顏鈺璇(Yu-Hsuan Yen)

指導教授 ： 陳弘森

高雄醫學大學/口腔醫學院/口腔衛生科學研究所碩士班/碩士(2012年)

https://doi.org/10.6832/KMU.2012.00101

摘要

背景
在2010年，口腔癌已排台灣十大癌症死因的第五名，而在男性族群癌症死因別排名更爬升到第四位，口腔癌儼然已成為一個重要的健康議題。基質金屬蛋白酶(MMPs) 和基質金屬蛋白酶之抑制劑(TIMPs) 參與了細胞外基質(ECM) 的降解、組織修復以及腫瘤細胞的發展。許多研究提到MMPs和TIMPs基因會參與癌症的發展過程，其中有報告指出MMP-13與TIMP-1基因在口腔癌組織中有異常表現的情形，而基因的表現可能會受到基因多型性(SNPs) 的調控，不過MMP-13和TIMP-1基因多型性是否參與口腔癌的發展仍然不清楚。

研究目的
本研究欲分析MMP-13 -77A>G以及TIMP-1 +372T>C 基因多型性對口腔癌與口腔癌前病變發生的風險與口腔癌前病變癌化潛力。

研究方法
本研究設計為病例對照法(case and control study) ，我們利用標準化問卷收集受試者的基本資料與物質(菸、檳榔、酒) 使用歷史以及抽取其周邊血液來萃取DNA，隨後利用TaqMan 探針與real-time PCR的方式判斷受試者的MMP-13 -77A>G與TIMP-1 +372T>C基因
型，並且使用PCR-RFLP與PCR產物進行核酸序列定序等方法來驗證。

結果
在使用物質狀態的分佈結果中，我們發現口腔癌以及口腔癌前病變患者有抽菸、喝酒、嚼食檳榔的盛行率相較於健康對照組非常的高。多變項邏輯式迴歸分析發現MMP-13 -77A>G基因多型性與口腔癌和癌前病變的發生或是口腔癌前病變癌化潛力並無顯著相關，進一步用不同物質使用習慣作分層分析，發現口腔癌的發生或口腔癌前病變癌化潛力與MMP-13 -77A>G基因多型性的相關性會因為有無嚼食檳榔或有無吸菸而不同。我們也發現女性族群的TIMP-1 +372C/C基因型與發生口腔癌有顯著相關；在沒有抽菸與沒有嚼食檳榔習慣的男性中，帶TIMP-1 +372C allele者與口腔癌的發生有關，有飲酒習慣且帶TIMP-1 +372C allele的男性罹患口腔癌前病變的風險相對於帶T allele者也較高。

結論
我們建議在女性族群，TIMP-1 +372C/C基因型可能是口腔鱗狀細胞癌的危險因子，而MMP-13 -77A>G基因多型性與口腔癌的發生沒有直接相關。我們也發現MMP-13 -77A>G與TIMP-1 +372T>C 基因多型性和菸、檳榔、酒等物質有交互影響的作用存在，進而參與口腔癌和口腔癌前病變的發生，以及口腔癌前病變癌化潛力的改變。

關鍵字

MMP-13 ； TIMP-1 ； 基因多型性 ； 口腔癌

並列摘要

Background
In 2010, mortality due to oral cancer in Taiwan was ranked as the fifth of the leading cause of cancer death and the fourth among males. Apparently, the prevention and control of oral cancer development has become an important health issue. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are involved in degradation of extracellular matrix (ECM), tissue remodeling and regulation in tumor cell progression. Many different MMP and TIMP genes were proposed to be involved in the cancer development, and the overexpression of MMP-13 and TIMP-1 genes was found in the oral cancerous tissue. The gene expression might be regulated by functional gene polymorphism, however, the effects of polymorphisms of MMP-13 and TIMP-1 genes on oral cancerous development were unclear.

Study objectives
Our aims were to investigate the risks of MMP-13 -77A>G polymorphism and TIMP-1 +372T>C polymorphism to the development of oral cancer and oral precancerous lesions and the malignance potential of oral precancerous lesions.

Methods
The case-control study was applied to determine the genetic susceptibility in this study. A standardized questionnaire was applied to collect the demographic data and histories of substances’ usage. Genomic DNA was extracted from peripheral blood samples. Polymorphisms in MMP-13-77A>G and TIMP-1 +372T>C genes were determined using TaqMan probe real-time PCR method. The results of genotyping were confirmed using the PCR-RFLP and PCR product direct sequencing methods to complete the genotype analysis for the 10% random selected samples.

Results
The prevalence of smokers, drinkers, and betel-quid chewers were significantly higher in OSCC and precancerous patients. Polymorphisms in MMP-13 -77A>G gene were not associated with development of OSCC and oral precancerous lesions (OPLs) and malignance potential of OPLs by multivariate logistic regression analysis. Stratification analysis with different substance’s usage showed that the risks of OSCC development and malignance potential could be differentiated by betel quid chewing and cigarette smoking. In female group, TIMP-1 +372C/C genotype was significantly associated with OSCC development. In male population, TIMP-1 +372C allele was associated with development of OSCC in non-smokers and non-betel quid chewers and OPLs in drinkers.

Conclusion
We suggested that TIMP-1 +372C/C genotype may be the risk factor for OSCC development in female. MMP-13 -77A>G polymorphisms were not associated with OSCC development. The effect of gene-environment interaction between MMP-13 -77A>G and TIMP-1 +372T>C polymorphisms and different substance’s usage on the development of OSCC and OPLs and malignance potential of OPLs were found in this study.

並列關鍵字

MMP-13 ； TIMP-1 ； Single nucleotide polymorphism ； Oral cancer

參考文獻

參考文獻

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