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Title: | 口腔鏈球菌與口腔鱗狀細胞癌的關係與交互作用 Association and interaction of oral streptococci with oral squamous cell carcinoma. |
Authors: | Wen-Hsiu Chen 陳玟秀 |
Advisor: | 賈景山(Jean-San Chia) |
Keyword: | 口腔癌,口腔鏈球菌, oral cancer,oral streptococci, |
Publication Year : | 2007 |
Degree: | 碩士 |
Abstract: | [背景和目的]:癌症與癌症組織內所含有細菌叢的關係是一重要研究方向,根據流行病學及以實驗室為基礎的研究都指出某些種類細菌與不同類型癌症有關,但詳細的分子機制仍未清楚。最早被提出也探討最多的是幽門螺旋桿菌,它會導致胃潰瘍,胃黏膜相關淋巴组織淋巴瘤和胃腺癌的產生。此外其他慢性感染細菌如傷寒桿菌與膽囊癌有強烈流行病學上的相關性、嗜檸檬酸桿菌感染實驗鼠引起大腸息肉增生與大腸癌有關、砂眼披衣菌感染會增加子宮頸癌發生的機率、牛鏈球菌則與大腸的惡性腫瘤有關等。至於口腔癌和細菌的關係也有被提出,例如咽峽炎鏈球菌、輕鏈球菌等。目前口腔及咽喉癌居世界最常發生之惡性腫瘤排名第六位,在發展中國家如東南亞地區,約40%的惡性腫瘤是位於口腔,本國行政院衛生署統計口腔癌的發生率和死亡率有逐年增加的趨勢,94年度公佈台灣地區十大常見癌症排名中,口腔癌已大幅晉升為第6名,在男性癌症中死亡率與發生率皆位居第四位,可見此等癌症已慢慢威脅到本國人民之健康,其嚴重性可想而知。由於目前提出口腔癌與特定細菌叢關係的文獻不多,因此本實驗的目的是針對已被提出與口腔癌有關的菌種及其他口腔鏈球菌屬,作進一步的探討,包括以下幾點:(1) 確認口腔鱗狀細胞癌組織中特異性菌種的分佈 (2)比較口腔鱗狀細胞癌患者及非口腔鱗狀細胞癌患者的唾液中,特異性菌種分佈的不同。(3) 以特異性菌種剌激口腔癌細胞株後所產生的變化。
[方法]:我們總共收集了38個口腔鱗狀細胞癌組織、29個鄰近腫瘤的正常組織、10個非口腔癌患者口內正常牙齦組織、5個卵巢癌組織及2個大腸癌組織,另外還有38個口腔鱗狀細胞癌患者的唾液樣本及38個非口腔鱗狀細胞癌患者的唾液樣本。實驗方式是將樣本的DNA萃取出來,利用PCR擴增出細菌16s rDNA基因片段,再以逆向棋盤式雜交分析樣本中細菌種類。另外利用化學冷光方法來測量菌種剌激細胞株後產生反應性含氧物種的量。 [結果]:實驗結果發現口腔鱗狀細胞癌組織中特異性菌種的分佈與其他組織不同,其中以輕鏈球菌的差異最大。口腔鱗狀細胞癌患者及非口腔鱗狀細胞癌患者的唾液中,特異性菌種的分佈則無明顯差異。我們同時證明輕鏈球菌剌激口腔癌細胞株之後會使受剌激的細胞明顯產生反應性含氧物種,這暗示輕鏈球菌與破壞DNA有關。 [Background and purpose]: The association of bacteria with cancer is an important issue for investigation. A number of bacterial species have been associated with different cancers by either epidemiological or laboratory-based studies. However, the underlying cellular mechanisms are far from clear. Helicobacter pylori infection has been considered to be causative agent of peptic ulcer, mucosa-associated lymphoid tissue lymphomas and gastric adenocarcinoma. Chronic infection with other bacteria, notably Salmonella typhi, is associated with gallbladder carcinoma. Citrobacter rodentium infection in mice causes a colonic hyperplastic disease that can lead to colorectal cancer. Chlamydia trachomatis infection has been associated with an increasing risk for the development of invasive cervical carcinoma. Streptococcus bovis have been associated with malignancies in the colon, while Streptococcus anginosus and Streptococcus mitis, were associated with oral cancer. At present, oral cancer is the sixth most common malignancy worldwide and is particularly prevalent in developing countries, such as in Southeast Asia. In Taiwan, oral caner ranked as the sixth most common cancer in both sexes and account for the fourth most common cancer in male population, with increasing incidence. We investigated the association of the specific bacterial flora including oral streptococci with the oral squamous cell carcinoma(OSCC). Our aim was to (1) confirme the identity as well as distribution frequency of specific bacterial flora within oral squamous cell carcinoma (2) compare the different distribution of specific bacterial flora between saliva from subjects with or without OSCC (3) investigate the interaction of specific bacteria with oral carcinoma cell line in vitro. [Method]: We collected samples from 38 oral squamous cell carcinoma and 29 their adjacent corresponding normal tissues from 10 gingival tissue of non-cancer patients, from 5 ovarian cancer and from 2 colon cancers We also collected saliva sample from 39 patients with OSCC and another 39 patients without OSCC. The DNA fragments encoded for 16S rRNA gene of DNA isolated from samples were amplified using universal primers. The amplicons were hybridized to the capture probes by checkerboard format to determine the bacterial species in samples. In addiction, stimulating oral cancer cells with specific bacteria and then reactive oxygen species production in cells were detected using a chemiluminescence technique. [Result]: A significant difference in the bacterial diversity, especially Streptococcus mitis, was found between oral squamous cell carcinoma and normal tissue. But no significant difference could be found in the contribution of bacterial flora in saliva between subjects with and without OSCC lesion. We also demonstrated that Streptococcus mitis could stimulate the ROS production dose-dependently in oral cancer cell line. This result implicated that Streptococcus mitis may be related to the DNA damage frequently encountered during carcinogenesis。 |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25069 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 臨床牙醫學研究所 |
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