口腔黏膜下纖維化是口腔黏膜慢性且持續性纖維化的癌前病變，口腔黏膜長期處在區域癌化的環境，使得惡性轉變更難以預測。過往針對口腔癌的檢體和正常組織相比，發現有特定細胞激素表現量不同。在大型的全國口腔黏膜篩檢的分析中，高危險族群日後轉變成口腔癌的唾液中亦有特定蛋白質高度表現。因此我們假定唾液中存有的細胞激素可能扮演著促進惡性轉變的功能，另外癌前病變組織或是惡性細胞的自分泌或是旁分泌信號也可能促進癌化。
首先我們分析了來自成大醫院口腔外科部門的 12 個臨床檢體，其中 6 個是單純患有口腔黏膜下纖維化的患者(並沒有其他癌前病變)，另外 6 個則是除了口腔黏膜下纖維化還併有口腔癌的患者，使用人類細胞激素晶片分析，結果呈現在有口腔癌的組別，其細胞激素 CXCL 1 以及 macrophage inhibitory factor (MIF)的表現量都有明顯增加。
接著使用柳營奇美醫院自2012-2017的臨床唾液檢體來驗證這兩種細胞激素，利用酵素免疫分析法(ELISA)定量 CXCL 1 和 MIF，在口腔癌組別和單純口腔黏膜下纖維化的組別相比，CXCL 1 和 MIF 表現量在口腔癌組別都有較高的趨勢。分析 CXCL 1 和 MIF 表現量與臨床危險因素的相關性，在口腔癌細胞分化部分，CXCL 1 的濃度在重度分化和輕度分化的組別有統計上的顯著差別，在重度分化的組別，具有較高濃度的 CXCL1。
非侵入性診斷與預後是癌症精準醫療的趨勢，透過唾液細胞激素的檢測未來或可發展為口腔癌前病變惡轉為口腔癌非侵入性診斷與預後的指標，將有助於口腔癌的預防與治療。

Oral submucous fibrosis is a chronic and progressive fibrosis lesion of the oral mucosa and is considered as a precancerous lesion. The oral mucosa has been in a field cancerization for a long time, making the malignant transformation more difficult to predict. In the past, specimens for oral cancer were found to show different
amounts of specific cytokines compared to normal tissues. In a large-scale national analysis of oral screening, specific proteins are also highly expressed in saliva from high-risk populations that later turn into oral cancer. Therefore, we hypothesized that cytokines in saliva may play a role in promoting malignant transformation. In addition, autocrine or paracrine signals from precancerous lesions or malignant cells may also promote carcinogenesis.
First, we analyzed 12 clinical specimens from the department of oral and maxillofacial surgery of Cheng Kung University Hospital, of which 6 were OSF patients , and the other 6 were patients with OSF combined with OSCC were analyzed by human cytokine array. The results showed that in the group , OSF combined with OSCC , the cytokine level of CXCL 1 and macrophage inhibitory factor (MIF) were significantly increased.
Then we used saliva samples collected from department of oral and maxillofacial surgery Chi Mei Hospital, Liouying campus from 2012-2017 to verify these two cytokines, and use ELISA to quantify CXCL 1 and MIF. Compared with the group,
the expression of CXCL 1 and MIF had a higher trend in the OSF combined with OSCC group. To analyze the correlation between the expression of CXCL 1 and MIF and clinical risk factors. The concentration of CXCL 1 has statistically significant difference in the poor differentiated group.
Non-invasive diagnosis and prognosis tools are the trend of precision medicine. The salivary cytokines may be developed into a detection tool that point out oral precancerous lesion become a cancerous lesion, which will help prevent and treat oral cancer.

中文摘要..…………………………………………………………….…………………... II
Abstract….………………………………………………………………..………………III
致謝………………………………………………………………………………………...V
Contents………………………………...………………………………………………...VI
Introduction………………………………………………………………………………..1
Oral cancer ……………………………………………………….……………………..1
Oral submucous fibrosis .….……………………………………….…………………...1
Field of cancerization…………………………………………………..………………..3
Salivary biomarker…………………………………………………….………………..4
CXCL1………………………………………………………………….………………..5
Macrophage migration inhibitory factor (MIF) …………………….………………..6
Motivation………………………………………………………………………………….8
Materials and methods .…………………….……………………………………………..9
Sample collection …………………….……………….…………………………………9
Saliva collection …………………….…………………………………………………...9
Cytokine array…………………….…………………………………………………...10
Protein expression assay……..………….……………………………………………..11
Enzyme-Linked ImmnuoSorbent Assay, ELISA…………………………………….11
Statistical analysis…………………….………………………………………………..13
Results……………...…………………….………………………………………………..14
1. Selection of overexpressed cytokines in saliva from OSF-OSCC patients by
cytokine array analysis…………...……...……………………………………………14
2. Quantification of cytokines levels in salivary samples of OSF patients and OSF-
OSCC patients….……………………………………………………………………15
3. Analysis of correlation of CXCL1 and MIF………………………………………15
4. Retrospective analysis the correlation between clinicopathological
characteristic and cytokines level…………………………..…………………………15
Discussion ……………………………………………………………………..………… 17
Conclusion……………………………………………………………………………...…21
References……………………………………………………………….………………..22
Tables and figures…………………………………………………….…………………..26
Table 1. Clinicopathological characteristics of OSF patients and OSF-OSCC
patients……………………….…………………….…………….……………….……26
Table 2. Clinicopathological characteristics (risk factors) of OSF-OSCC patients.. 27
Figure 1. Human cytokine arrays showed the levels of CXCL1 and MIF are higher in saliva from OSF-OSCC patients than those from OSF patients……………...…29
Figure 2. Quantification of cytokine concentration in saliva………………………...30
Figure 3. The correlation between cytokine concentration and age ＆ MMO in all
patients……………………………….…………………….…………………………..31
Figure 4. Comparison of the correlation between CXCL1 level and clinicopathologic
factors in OSF combined with OSCC patients………………… ..…………………32
Figure 5. Comparison of the correlation between MIF level and clinicopathologic
factors in OSF combined with OSCC patients………..………………………………33

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