臺灣博碩士論文加值系統

口腔鱗狀細胞癌是世界上常見惡性腫瘤之一，根據中華民國行政院衛生福利部國民健康署105年統計數據，口腔癌在台灣的死亡率仍不斷的增加，即使現今醫療進步，對口腔鱗狀細胞癌的治療已經有明顯的進展，但預後不良的狀況仍困擾著病患，而科學家們為了追求更好的療效積極開發新藥。喹唑啉酮衍生物在30多年前就已被運用於抗癌，包含血癌、肝癌、肺癌、口腔癌等。在此研究中，我們使用新合成喹唑啉酮衍生物 (HoLu 1-16) 來研究其對鱗狀口腔癌細胞的抗腫瘤作用。利用細胞存活實驗，我們發現HoLu-12可以顯著的抑制口腔癌細胞的存活率，細胞週期分析結果顯示，HoLu-12會使口腔癌細胞停滯在G2/M時期，並使sub G1時期的細胞數增加。西方墨點法結果發現HoLu-12會造成細胞凋亡蛋白的表現量增加。動物實驗結果發現，HoLu-12會明顯抑制小鼠體內的腫瘤生長。此外，HoLu-12與5-Fu的合併使用呈現協同作用。這些結果表明 HoLu-12具有針對口腔癌細胞的治療潛力。

Oral squamous cell carcinoma (OSCC) is one of the common malignant diseases in the world. Through advances in multimodality therapies for the treatment of OSCC; however, the overall 5-year survival rate for OSCC patients remains at 50% without significant improvement over the past three decades. Therefore, it is necessary to develop novel potential therapeutic agents against human OSCC. Quinazolinone is a heterocyclic chemical compound having a unique place in the field of medicinal chemistry. Many quinazolinone derivatives possess a wide range of bioactivities such as antimicrobial, anti-inflammatory, anticonvulsant, analgesic, anthelminthic and anticancer activities. In this study, we examined the therapeutic potential of 16 synthetic quinazolinone derivatives in OSCC by the sulforhodamine B (SRB) cytotoxicity assay. Among them, HoLu-12 exhibited no cytotoxicity toward the human normal oral mucosa fibroblast (OMF) cells. However, HoLu-12 significantly reduced the viability of several human OSCC cell lines, such as CAL27 and OECM-1. Cell cycle analysis revealed that HoLu-12 induced G2/M arrest and sub G1 phase in OSCC cells. Western blot analysis showed that treatment with HoLu-12 increased the expression of cleaved PARP, indicating that HoLu-12 could induce apoptosis pathway. Subcutaneous xenograft model was performed to demonstrate that HoLu-12 could significantly inhibit OSCC growth in vivo with negligible body weight loss. In addition, we observed that combined treatment with HoLu-12 and 5-flurouracil had a synergistic cytotoxic effect in OSCC cells. Taken together, these results suggest that HoLu-12 may have the therapeutic potential against human OSCC cells.

目錄
壹、 中文摘要---------------------------------------------------------- 6
貳、 英文摘要---------------------------------------------------------- 7
參、 研究背景---------------------------------------------------------- 9
●口腔癌簡介----------------------------------------------------------- 9
●細胞癌化與死亡機轉----------------------------------------------- 11
●喹唑啉酮 (Quinazolinone) 的背景、衍生物與抗癌研究---------------------------------------------------------------------------- 13
肆、 研究動機與目的--------------------------------------------- 15
伍、 實驗材料與方法--------------------------------------------- 16
一、 材料----------------------------------------------------- 16
二、 實驗方法----------------------------------------------- 18
1. 細胞培養、計數------------------------------------- 18
2. 細胞存活率測試-------------------------------------- 19
3. 乳酸脫氫酶實驗-------------------------------------- 19
4. 動物實驗----------------------------------------------- 20
5. 細胞週期分析----------------------------------------- 21
6. 西方墨點法------------------------------------------- 22
7. Apoptosis antibody array---------------------------- 23
8. 藥物合併指數-----------------------------------------24
9. 實驗數據統計分析----------------------------------- 24
陸、 實驗結果----------------------------------------------------------- 26
一、 探討16種喹唑啉酮衍生物對口腔癌細胞的細胞毒殺作用------------------------------------------------------------------------ 26
二、 偵測HoLu-12對口腔癌細胞的細胞壞死作用--------------- 26
三、 偵測HoLu-12對多株口腔癌細胞之IC50---------------------- 27
四、 測試HoLu-12在動物實驗中對口腔癌腫瘤的影響--------- 27
五、 觀察HoLu-12對口腔癌細胞的細胞週期影響--------------- 28
六、 觀察HoLu-12對口腔癌細胞的細胞凋亡影響----------------29
七、 確認HoLu-12所引起口腔癌細胞之細胞凋亡相關蛋白變化-------------------------------------------------------------------------29
八、 HoLu-12與5-Fu合併使用後對口腔癌細胞之毒殺效果----31
柒、 討論------------------------------------------------------------------- 33
捌、 結論------------------------------------------------------------------- 38
玖、 參考文獻---------------------------------------------------------- -39

表格目錄
Table 1. 喹唑啉酮衍生物HoLu-12 (C18H16FN3O2) 結構--------------- 44

圖目錄
Figure 1. 探討16種喹唑啉酮衍生物對口腔癌細胞的細胞毒殺作用------------------------------------------------------------------------------------- 45
Figure 2. 偵測HoLu-12對口腔癌細胞的細胞壞死作用---------------- 48
Figure 3. 偵測HoLu-12對多株口腔癌細胞之IC50---------------------- 49
Figure 4. 測試HoLu-12在動物實驗中對口腔癌腫瘤的影響---------- 50
Figure 5. 觀察HoLu-12對口腔癌細胞的細胞週期影響---------------- 52
Figure 6. 觀察HoLu-12對口腔癌細胞的細胞凋亡影響---------------- 55
Figure 7. 確認HoLu-12所引起口腔癌細胞之細胞凋亡相關蛋白變化------------------------------------------------------------------------------------- 57
Figure 8. HoLu-12與5-Fu合併使用後對口腔癌細胞之毒殺效果------------------------------------------------------------------------------------- 59

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