基質金屬蛋白酶14之基因多型性與肝細胞癌之相關性探討

Association of MMP-14 gene polymorphisms with the susceptibility and serverity of heptocellular carcinoma

洪幸珊(Hsing-Shan Hung)

指導教授 ： 謝易修

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2010年)

摘要

肝細胞癌 (Hepatocellular carcinoma；HCC) 簡稱肝癌，是一種源自肝細胞的惡性上皮性腫瘤，常見於慢性肝炎和肝纖維化的末期併發症，為目前全球排名第五名的惡性腫瘤，居所有癌症死亡率的第三名。造成肝癌的真正成因至今尚未十分明瞭，根據研究指出受到環境和遺傳因素的共同影響。肝癌具有高度轉移、再復發及預後不佳等特性。基質金屬蛋白酶 (Matrix metalloproteinase；MMP) 是一群含鋅金屬離子的細胞外基質水解酶，它們透過對細胞外基質造成不同程度的降解，在腫瘤的侵襲與轉移過程扮演重要的角色。過去的研究顯示在肝癌組織中MMP-14 有大量表現的情形，MMP-14不僅促進肝癌細胞的生長、侵襲及轉移並且與患者預後不佳有關。MMP-14 是一種膜型金屬蛋白水解酶，與細胞的多種生理及病理過程有關。MMP-14 具有降解細胞外基質成份、水解細胞表面蛋白與活化 pro-MMP-2 的作用，對於細胞功能與微環境的修飾以及細胞外基質的重塑扮演重要的角色。文獻指出金屬蛋白水解酶的基因多型性會透過影響基因的轉錄和蛋白的表達而與不同腫瘤的易感性、疾病嚴重程度及預後有關。本研究的目的在於探討 MMP-14 之基因多型性是否與肝癌的罹患敏感性有關，以及是否與肝癌病患的疾病嚴重程度有相關性。本研究一共收集了 102 位肝癌患者及 347 位正常成人作為對照組，分別進行 MMP-14 -165 G/T 、+7096 T/C 與 +8153 G/A 之基因多型性分析。實驗組與對照組的 MMP-14 基因多型性測定是以聚合酶連鎖反應與限制酶片段分析基因多型性方法來偵測，並以統計分析方法來判定兩組之間的差異性。同時也分析 MMP-14 之基因多型性與肝癌相關臨床參數，包括臨床分期、腫瘤大小、淋巴結轉移、遠端轉移、肝功能指標、B 型肝炎抗原、C 型肝炎及肝硬化等的關係。結果發現 MMP-14 ＋7096 位置上的變異在肝癌病例與對照組之間基因頻率的分佈具有顯著差異 (p = 0.019)，攜帶 T 對偶基因罹患肝癌的比例較攜帶 C 對偶基因罹患肝癌的比例高 (p = 0.005)。在 MMP-14 -165 基因方面，攜帶 G/G 基因型的肝癌患者相較於攜帶 G/T 或 T/T 基因型的肝癌患者，明顯的肝臟機能比較差 (p = 0.005)。因此我們認為，MMP-14 之基因多型性與中台灣族群形成肝癌的罹患敏感性有關，並且也與肝癌病患的疾病嚴重程度有相關性。

關鍵字

肝細胞癌 ； 基質金屬蛋白酶

並列摘要

Hepatocellular carcinoma (HCC) is an epithelial cancer that originates from hepatocytes and the most common primary malignant tumor of the liver. The majority of HCC arise in the chronically injured liver as a result of chronic hepatitis and liver cirrhosis. HCC is the fifth most common cancer worldwide and the third most common cause of cancer mortality. At present, the pathogenesis mechanisms of liver cancer are not entirely clear. Long-term prognosis of patients with HCC is generally poor, and survival is mainly affected by the occurrence of metastases. Matrix metalloproteinases (MMPs) are a broad family of zinc-binding endopeptidases that play a key role in the extracellular matrix (ECM) degradation associated with cancer cell invasion, metastasis and angiogenesis. Destruction of the ECM by MMPs has been shown to play an important role in the migration and spreading of tumor cells, leading to invasion and metastasis. Study showed that the MMP-14 gene is strongly expressed in hepatocellular carcinoma cells and is involved in the invasion potential of hepatocellular carcinoma. MMP-14 is a trans- membrane metalloprotease that plays a major role in the extracellular matrix remodeling, directly by degrading several of its components and indirectly by activating pro-MMP2. MMP-14 is indeed a micro- enviroment modifier and also a cell function modifier. Recently, functional gene polymorphisms in MMPs have been found, and some reports have revealed an association between these polymorphisms and the susceptibility of various cancers. The aim of the present study was to investigate the association of MMP-14 gene polymorphisms (-165 G/T, +7096 T/C and +8153 G/A) with the susceptibility and serverity of heptocellular carcinoma. A total of 102 patients with HCC cases were obtained from Chung Shan Medical University Hospital. Of which 347 control subjects were selected from people who came for routine physical checkups. Polymorphism of sites within the promoter region and exon of MMP-14 gene were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique on genomic DNA extracted from whole blood. Statistical analysis were conducted to explore the differences in the distribution of alleles and genotypes between patients and controls. But also analysis of MMP-14 gene polymorphisms and clinical features of liver cancer, including clinical stage, tumor size, lymph node metastasis, distant metastasis, Child-Pugh grade, HBsAg, AntiHCV and cirrhosis. Result indicated that there was a significant difference in the distribution frequency of MMP-14 +7096 polymorphism between case and control group (p = 0.019), carry T allele increased the susceptibility to HCC than carry C allele (p = 0.005). In HCC clinical features, individuals with -165 G/G genotype had a poor liver synthetic capacity as compared to those with G/T or T/T genotype in the HCC patient group (p = 0.005). These results have demonstrated a significant association between the polymorphisms of MMP-14 gene with the susceptibility and serverity of HCC in the Taiwanese population.

並列關鍵字

Hepatocellular carcinoma ； Matrix metalloproteinases

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