嚼食檳榔的口腔癌患者中第一型基質金屬蛋白酶的表現

Expression of Matrix Metalloproteinase-1 in Oral Cancer Patients from Betel Quid Users

邱昶達(Chang-Ta Chiu)

指導教授 ： 劉巡宇

臺北醫學大學/口腔醫學院/牙醫學系碩博士班/碩士(2008年)

https://doi.org/10.6831/TMU.2008.00121

摘要

口腔癌中以鱗狀細胞癌 (squamous cell carcinoma) 所佔的比率最高，比率高達 95%，且有八至九成的患者是長期的檳榔使用者。檳榔中各種成分會對口腔粘膜產生機械與化學性傷害，許多研究已證實檳榔嚼食者的粘膜常會出現上皮萎縮或過度角化增生，甚至引發口腔癌。基質金屬蛋白酶 (matrix metalloproteinases，MMPs) 是一群鋅離子依賴型 (Zinc dependent) 的內胜肽酶 (endopeptidase)，參與了細胞外基質的生理性降解過程，但是 MMPs 同時也影響了一些疾病的病理過程。在腫瘤侵犯過程中，上皮細胞轉移進入下方的結締組織是一個重要特徵，此作用伴隨著細胞與基質間交互作用的改變和基質降解酶的分泌，其中最重要的就是 MMPs。在腫瘤細胞的成長與增生中，細胞外基質間 MMPs 的調控是很重要的， MMPs 會重塑腫瘤細胞周圍的組織；在腫瘤細胞、細胞外基質、發炎細胞間的相互作用被認為會誘導 MMPs 的表現，因此了解腫瘤細胞中 MMPs 的高低表現，對預測腫瘤細胞的侵入與轉移是十分有意義的。其中，第一型基質金屬蛋白酶 (MMP-1) 又稱為纖維母細胞膠原蛋白酶，主要功能在裂解結締組織中第一、二、三型膠原蛋白， MMP-1 的高度表現已被證實與大腸直腸癌、肺癌、食道癌有關，近來更陸續有研究指出 MMP-1 的高度表現與口腔癌發生、口腔癌增生、淋巴轉移及其預後有關。特別是在口腔腫瘤中，分解口腔黏膜下結締組織中含量最多的細胞外基質，對於口腔鱗狀細胞癌的進行是很重要的，因此參與降解細胞外基質有關的 MMP-1 被認為可以成為口腔腫瘤細胞標記與預測腫瘤的預後。但是目前已發表的 MMP-1 相關研究中屬於國外口腔腫瘤患者為主，缺乏專門針對多有嚼食檳榔習慣的國內患者，因此本研究計劃針對嚼食檳榔習慣盛行的國內口腔癌患者，分析台灣口腔癌患者中 MMP-1 於組織中的分布情況。共選取了奇美醫學中心口腔顎面外科 30 位只具有長期嚼食檳榔並接受口腔癌手術切除的患者，分別進行腫瘤樣本組織與腫瘤周圍組織中 MMP-1 的半定量反轉錄作用-聚合酶連鎖反應 (RT-PCR)，發現在腫瘤樣本組織中 MMP-1 mRNA 有普遍提高的表現。此外，為了評估腫瘤組織中 MMP-1 蛋白質分子的表現，選取口腔腫瘤手術切除患者的腫瘤組織與腫瘤周圍組織，以西方墨點法分析確認在腫瘤組織中 MMP-1 蛋白質的含量提高表現，同時選取口腔癌手術切除患者的腫瘤樣本組織，進行組織免疫化學染色法 (immunohistochemical staining，IHC) 分析，發現在腫瘤上皮組織中 MMP-1 也提高表現向。再者， 30 位患者皆具有長期嚼食檳榔的習慣，因此我們藉由模擬在口腔癌細胞株與食道癌細胞株中，觀察細胞培養基接受檳榔嚼塊萃取液與檳榔素的篩選刺激後，發現經由檳榔成份篩選刺激後的腫瘤細胞，會提高 MMP-1 的表現。以上結果可以證明， MMP-1 對於嚼食檳榔的口腔癌患者的腫瘤發生、腫瘤增生、淋巴結轉移與腫瘤分化應該扮演一定的角色。

關鍵字

口腔癌 ； 第一型基質金屬蛋白酶檳榔

並列摘要

The squamous cell carcinoma (SCC) rate is the highest (95%) among oral cancers. About 80-90% of the patients of oral cancer chew betel nuts for a long time. Most ingredients of the nuts may hurt the oral mucosa mechanically and chemically. Betel nut chewing has been proved to cause epithelial atrophy, hyperkeratosis, even oral cancer. Matrix metallo- proteinases (MMPs), a group of zinc-dependent endopeptidases, involve extracellular matrix (ECM) physiological degradation process. Moreover, MMPs influence other pathological processes of diseases. During the tumor invasion, cancer cells break through the basement membrane and enter connective tissue, which is a crucial basic characteristic, and varies with the changes in the interaction of cells, matrix and matrix degrading enzymes. Among the enzymes, MMPs is one of the most important. During tumor cell proliferation, the adjustment between extracelluar matrix and MMPs is important. MMPs can remodel tumor cells of peripheral tissue and be induced in the expression during the interaction among tumor cell, ECM, and inflammatory cell. Thus MMPs are considerably important in tumor tissue remodeling, repairing and destroying. Thus the up- and down- expression of MMPs is significantly different when applied to predict tumor cell invasion and transfer. The overexpression of some MMPs were associated with oral cancer occurrence, proliferation, lymph node metastasis and prognosis in recent studies.

Matrix metalloprotinase-1 (MMP-1) cleave type I, II and III collagen in connective tissues. The overexpression of MMP-1 is associated with oral cancer occurrence, proliferation, lymph node metastasis and prognosis in recent studies. MMP-1 is considerably important in degrading of ECM. Thus the up- and down- expression of MMP-1 is significantly different when applied to predict tumor invasion and prognosis. But the overseas MMP-1-associated cases of oral cancer lacked those of Taiwan, where there are many betel nut chewers. Therefore, this research aimed at Taiwan, to analyze the MMP-1 expression of oral cancer in the just- mentioned patients. The Department of Oral and Maxillofacial Surgery of Chi-Mei Medical Center especially studies 30 oral cancer patients of betel nut chewing and about the associated MMP-1 expression and relation. In RT-PCR analysis, we found that MMP-1mRNA was overexpressed in all the tested 30 OSCC. In Western blotting analysis, levels of MMP-1 protein was significantly elevated in the tested OSCC specimens than in their adjacent tissues. In IHC analysis, MMP-1 expression is also higher in the epithelium of tumor tissues. Esophageal carcinoma (CE81T/VGH) and OSCC (OECM-1) cell lines survived from the cytotoxic BQ extract (BQE) and arecoline selection process were found to express higher MMP-1 mRNA. These reports proved the significant effect of MMP-1 in betel nut chewing associated oral cancer, tumor growth, tumor differentiation and lymph node metastasis.

並列關鍵字

Oral cancer ； Metalloproteinase-1 ； Betel nut.

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