- 學位論文
KLK7與KLK10基因多型性與口腔癌之相關性分析
Genetic Polymorphisms of KLK7 and KLK10 and oral cancer risks
摘要
背景 根據衛生署民國九十八年統計，口腔癌在台灣十大癌症中死亡率位居第六位，且有逐年增加的趨勢。於2008年與日本大學松戶齒學部合作利用Affymetrix GeneChip Human Genome U133 Plus 2.0 Array 分析，探討口腔鳞狀上皮細胞癌組織與遠端safety margin組織的基因表現的差異，發現KLK7與KLK10基因有明顯較高mRNA表現量，而且是在血液中可以偵測得到的蛋白質。過去研究顯示，KLK7與KLK10基因可能與許多不同的癌症有關，但與口腔鳞狀上皮細胞癌的相關文獻並不充分。因此本研究將針對KLK7與KLK10基因多型性與口腔癌的相關性進行探討。 研究目的 我們將探討KLK7和KLK10基因多型性與口腔癌發生間的相關性。並進一步討論這兩個基因多型性與癌前病變的發生和口腔癌前病變惡化潛力的相關性。 研究方法 研究對象皆為男性且有十年以上的嚼食檳榔習慣，口腔癌前病變患者95名及口腔鱗狀上皮癌患者264名則是從高雄醫學大學附設醫院口腔外科邀請其自願參加，健康對照組為無口腔黏膜病變的人共173名。KLK7和KLK10基因上的基因多型性（rs10581213, and rs3745535, respectively）之決定是以聚合酶鏈反應 (Polymerase Chain Reaction) 將KLK7基因與KLK10基因片段複製，加上具有基因多型性位置具有特異性螢光標記的探針，再使用ABI Prism 7500進行最終PCR產物的螢光偵測（Applied Biosystems; Foster City, CA, USA）。 結果 KLK7與KLK10基因多型性與癌前病變的發生和口腔癌前病變的惡化潛力無顯著相關。在嚼食檳榔的族群中，KLK10基因多型性和口腔癌有顯著關係，有GT +TT基因型的人得到口腔癌的風險較高（AOR = 2.00; 95%CI = 1.31–3.09）。KLK7基因中帶有AACC insertion 同合子與異合子在未調整可能的干擾因子時可觀察到增加了口腔癌的風險，但調整可能的干擾因子後並未發現有顯著差異。而在菸、酒、檳榔習慣都有的族群中， KLK10基因帶有GT與 TT基因型有1.78倍的危險得到口腔癌，且在調整其他干擾因子後其顯著相關性依然存在，而KLK7基因多型性則未發現與疾病有顯著相關。 結論 本研究結果顯示在KLK7的基因多型性與口腔癌的發展並無顯著相關。在菸、酒、檳榔習慣都有的族群中，KLK10基因中帶有GT與 TT基因型有1.78倍較高的風險得到口腔癌。
並列摘要
Background According to Health Statistics of Department of Health in 2009, the mortality of oral cancer in Taiwan ranked sixth cancer in the entire population. In 2008, we cooperated with Nihon University School of Dentistry at Matsudo and identified that mRNA of KLK7 and KLK10 genes showed higher expression in OSCC tissues than normal tissues. Other studies also report that these two genes are associated with many cancers. However, the evidence exploring oral cancer is relatively few. Hence, we will evaluate the association between these two genes and oral cancer in this study. Objective In the present study, we examined whether the KLK7 and KLK10 polymorphisms is associated with oral cancer risk. Then, we also investigated the premalignant lesions risk and the malignant potential of oral premalignant lesions. Methods All study subjects are male and they were all betel quid chewers during over 10 years. In this study, we collected 95 oral premalignant patients and 264 OSCC patients from Kaohsiung Medical University Hospital. The control group has 173 people who has chewing habits but no oral mucosal lesions. We have genotyped two single nucleotide polymorphic sites in the KLK7 (rs10581213) and KLK10 (rs3745535) genes. Quantitative real-timePCR assays (rtPCR) were performed with gene-specific fluorescent labeled probes in a PCR ABI Prism 7500 Sequence Detector using TaqMan Universal PCR Master Mix (Applied Biosystems; Foster City, CA, USA). Results In this study showed that KLK7 and KLK10 polymorphisms were not correlated with premalignant development or malignant potential of premalignant. A significant association between oral cancer susceptibility and KLK10 polymorphism in betel quid chewers was demonstrated since individuals, have GT+TT genotypes, had a higher risk for oral cancer after adjusting for other confounders (AOR = 2.00; 95%CI = 1.31–3.09). We showed that the of AACC insertion homozygote plus AACC insertion heterozygotes in KLK7 gene increases the oral cancer risk but there was no significant difference. Moreover, results also revealed in KLK10 gene, the T/T plus G/T heterozygotes in cancer group were significantly more than control group and the odd ratio is 1.78 in betel quid chewer with drinking and smoking group, in KLK7 gene was not. Conclusion This is a case-control study which focuses on the polymorphisms of KLK7, KLK10 and oral cancer in Taiwanese male subjects with betel quid chewing habit. Results presented in this study showed AACC insertion homozygote plus of AACC insertion heterozygotes in KLK7 gene is not correlated with oral cancer development. In conclusion, the presence of the T/T plus G/T heterozygotes in KLK10 gene is associated with a higher risk of oral cancer.