根據最新台灣癌症登記資料和死因統計，近十年，台灣每年罹患口腔癌的人數已增加2倍，每年約有5,400名新診斷口腔癌個案，2,300人因口腔癌死亡。癌症之所以可怕，不在於腫瘤組織會不受控制地增殖，而在於它除了會在原發病灶附近造成破壞之外，更會產生遠端轉移，破壞肝、肺及腦等維生器官，因而造成病患死亡。過去文獻指出5-MTP這個內生性的色氨酸(tryptophan)的衍生物，可以抑制腫瘤的生長、降低腫瘤細胞轉移及減低發炎反應，進而得以殺死或控制癌細胞的生長。本論文研究在不同轉移能力的口腔癌細胞進行體內及體外的實驗。在細胞實驗方面在加入400μM 的5-MTP作用24小時後，口腔癌細胞運動、侵襲能力的降低。在動物實驗方面在5-MTP注射50天後，其腫瘤細胞的生長有被抑制的情形，並在肺臟的組織染色 (H&E)切片中也發現癌細胞在轉移後浸潤肺臟血管的比例也相對減低。而後再將細胞液的蛋白質樣品進行二維差異電泳 (2D-DIGE) 並結合基質輔助雷射脫附游離法 (MALDI-TOF MS) 進行分析。在蛋白質體學技術研究中我們鑑定出有65個蛋白質點有顯著性變化。而我們從中挑選4個蛋白(AKT、PGK1、TMP1及TMP4)來探討其對於降低口腔癌細胞的侵襲及轉移能力機制探討。發現PI3K/AKT/ IκB/NF-κB訊息傳遞路徑、PGK1訊息傳遞路徑及TMPs蛋白的表現量皆可由5-MTP色胺酸代謝物所調控，而抑制口腔癌細胞的存活及轉移的能力。這些表現量有差異的蛋白希望能提供未來治療和診斷的口腔癌指標。

粒線體在細胞中是一個很重要的胞器，它負責細胞的多種功能，包括能量代謝調節、細胞生長與分裂、自由基的產生並參與細胞凋亡路徑。曾有文獻研究指出，高糖刺激時，會導致粒線體的缺陷和氧化磷酸化及能量產生等機能失調。在我們的研究中，我們利用視網膜色素上皮細胞在不同糖濃度(5.5mM、50 mM及100 mM)來觀察三株細胞的存活率、細胞內過氧化物(ROS)的含量及粒線體膜電位(Δψm)的測試，得到在存活率方面三株細胞在24小時內細胞存活率沒有太大的變化，但在內生性的過氧化物及粒線體膜電位的測試中發現隨著糖濃度的增高兩者的含量也有隨之增高的情形。是從細胞中萃取粒線體以增加粒線體蛋白質的含量，再將粒線體蛋白質樣品進行二維差異電泳 (2D-DIGE) 並結合基質輔助雷射脫附游離法 (MALDI-TOF MS) 分析在不同糖濃度的視網膜色素上皮細胞之間有差異表現的粒線體蛋白質。在蛋白質體學技術研究中我們鑑定出有105個蛋白質點有顯著性變化，其中有12個蛋白質點是屬於粒線體蛋白質。我們發現高糖導致細胞內的ROS大量產生，進而調控粒線體中的VDAC1蛋白而誘導細胞釋放出發炎及凋亡的訊號分子；藉由調控TUFM蛋白的表現量而調控下游蛋白誘導細胞釋放出具有調控自我吞噬能力的蛋白。另一方面在不同糖濃度的視網膜色素上皮細胞中粒腺體膜電位的改變，而同時調控著電子傳遞鏈中的相關蛋白，如NDUFA2、UQCRC1、ATP5B及ATP5H等，促使氧化磷酸化無法正常的表現，進而產生更大量的過氧化物，造成粒線體的功能喪失。這些表現量有差異的粒線體蛋白及相關機制希望能提供未來在治療及診斷早期糖尿病眼病變的指標。

The metastasis status for oral cancer is highly associated with the overall survival rate of patients. Previous studies showed that the endogenous tryptophan metabolite, 5-methoxytryptophan (5-MTP) is able to down regulate COX-2 expression and to surpress tumor proliferation, migration and invasion as well as to reduce tumor size. To better understand the molecular mechanisms involved in the regulation of 5-MTP in the tumorogenesis of oral cancer, we conducted a proteomic analysis to identify the changes of protein expression as potential biomarkers to investigate the mechanism underlying cancer progression and metastasis. By using comparative wound healing assay and transwell invasion assay, our results revealed that 5-MTP can reduce invasive oral cancer cell migration and invasion ability. In SCID mouse, we discovered that primary tumor were inhibited by 5-MTP on both OC3 oral cancer and invasive OC3-I5 oral cancer cells. Moreover, enlarged spleen size in OC3-I5-induced SCID mouse has been observed, yet 5-MTP is able to inhibit this phenomenon. In the study of comparative proteomics, we have identified 65 protein spots showing significant changes by 2-DE and MALDI-TOF MS analysis. The differentially expressed proteins include Armadillo repeat-containing X-linked protein 1 (AKT), Phosphoglycerate kinase 1 (PGK1), Translin-associated protein X (TMP1) and Tropomyosin alpha-1 chain (TMP4), which have been reported to play potential roles in promoting cancer growth. We conclude 5-MTP plays a very critical role in inhibiting in vitro and in vivo cancer invasion and metastasis by targeting the signaling pathways of PI3K/AKT/ IκB/NF-κB、PGK1 and TMPs.

Retinal disease in patients with Type II diabetes is the leading cause of low vision and blindness, which is a major healthcare problem. Hence early identification of diabetic patients prone to develop this complication is important. Since mitochondrial oxidative stress plays a primary role to activate the intrinsic apoptotic cascade and trigger cellular damage, it is useful to utilize high glucose to induce excessive oxidative stress to study mitochondrial dysfunction and other early events of diabetic retinopathy. Herein, our data showed that reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) had been up-regulated in high glucose induced retinal pigmented epithelium cells (ARPE-19) . Furthermore, we conducted a mitochondrial proteomic analysis of retinal pigment epithelial cells with differently high-glucose concentrations. Our results showed that 105 protein spots with significant changes were identified by 2-DE and MALDI-TOF MS analysis in the comparative proteomics. Of the overall 105 proteins, 12 protein spots belong to known mitochondrial proteins (11%). The differentially expressed mitochondrial proteins include VDAC1, FH, TUFM, NDUFA2, UQCRC1, ATP5B, ATP5H, HSPD1, GOT2, MTRF1, SQRDL and PRDX3. All above proteins are able to regulat mitochondrial ATP synthesis, TCA cycle, Ca2+ storage and electron transfer chain (ETC). Accordingly, these mitochondria proteins cause ATP synthesis decline, oxidative stress increase and induce cell during pro-apoptosis or pro- inflammatory and more overexpressed ROS. All above mechanisms of mitochondrial proteins may help to understand how high blood sugar levels damage mitochondria and lead to the development of diabetic retinopathy, which will shed a light on the prevention of this serious diabetes-related eye disease.

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