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Investigations for targeting signaling pathways in the apoptosis induction and anti-metastasis of the novel quinazolinone derivatives in leukemia and oral cancer cells in vitro and anti-tumor activity in vivo
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|摘要:||先前的研究已證實顯示2-phenyl 6-pyrrolidinyl-4-quinazolinone衍生物具有抗癌活性和生物反應。其中，MJ-29與HMJ-38為二者最具潛力的新合成化合物，然而其抗癌活性機轉既不清楚也無得到很好的釐清。在本篇論文中，我們試圖探討在離體外和活體中抗血癌與口腔癌的作用與效果。在第一章中，我們著重於新穎化合物MJ-29在離體外骨髓單核細胞性小鼠血癌WEHI-3細胞中探討其對內質網壓力和粒線體依賴性細胞凋亡的影響，並推測MJ-29可能影響原位移植誘導的血癌小鼠。在第二章，其研究目標在探索MJ-29對於人類口腔鱗狀細胞癌CAL 27細胞的抗轉移作用的影響，並確認與發掘其相關分子機轉。在第三章中，探討另一新合成和具抗有絲分裂的化合物HMJ-38對於其標的與確切的機轉試驗，我們假設HMJ-38可能敏感化離體外CAL 27細胞的凋亡與其抑制裸小鼠活體上異種移植腫瘤的生長。在此研究中，我們證實發現在MJ-29處理的WEHI-3細胞離體試驗中藉由透過內在路徑與內質網壓力訊息傳遞誘導產生細胞凋亡現象；引人注目的是MJ-29抗血癌反應有在活體試驗血癌小鼠中被觀察到。接著，我們更發現MJ-29透過下調控基質金屬蛋白酶-2/-9（MMP-2/-9）在人類口腔鱗癌CAL 27細胞中能夠抑制細胞黏附、侵襲與移行；且磷酸化AKT和MAPK訊號路徑蛋白可協同參與涉及遭MJ-29抑制所影響的轉移必需步驟。最後，我們的研究提供HMJ-38所引起的CAL 27細胞G2/M期停滯與凋亡的證據與腹腔注射HMJ-38可抑制裸小鼠上CAL 27異種移植腫瘤的生長。總體而言，我們提出MJ-29 與HMJ-38擁有對抗血癌和口腔癌的潛力與治療價值的新發現和反應，再者二者化合物也許未來足以當作具有潛在的治療方式。|
The previous studies have demonstrated that 2-phenyl 6-pyrrolidinyl-4-quinazolinone derivatives exhibited antitumor actions and biological responses. Among them, both of the most potent newly synthesized compounds, 6-pyrrolidinyl-2-(2-hydroxyphenyl)-4-quinazolinone (MJ-29) and 6-pyrrolidinyl- 2-(3-Methoxyphenyl)-4-quinazolinone (HMJ-38) for their mechanisms underlying anticancer activities remain unclear and have not been well clarified. In this dissertation, we attempt to investigate anti-leukemia and anti-oral cancer effects in vitro and in vivo. In the chapter 1, we focused on the in vitro effects of MJ-29 on ER stress and mitochondria-dependent apoptotic death in murine myelomonocytic leukemia WEHI-3 cells, and to hypothesize that MJ-29 might fully impair the orthotopic leukemic mice. In the chapter 2, the objective of current study was explored the effects of the novel compound MJ-29 on anti-metastatic actions of human oral squamous cell carcinoma (OSCC) CAL 27 cells and to verify the underlying related molecular mechanisms in this event. In the chapter 3, this study investigated the newly synthesized and anti-mitotic compound, HMJ-38 addressing its target and precise mechanism of actions. We hypothesized that HMJ-38 might sensitize apoptotic death of CAL 27 cells in vitro and inhibit xenografts tumor growth in vivo. In my Ph.D. study, we found that the induction of cell death in MJ-29-treated WEHI-3 cells has been proven by an assessment of apoptosis in vitro culture through intrinsic apoptotic pathway and ER stress signaling. Strikingly, anti-leukemia responses by MJ-29 were observed in leukemic mice in vivo. We further demonstrated that MJ-29 is capable of inhibiting cell adhesion, invasion and migration through the down-regulation of MMP-2 and MMP-9 in human OSCC CAL 27 cells. Also, the phosphorylation of AKT and MAPK signaling pathway proteins may be coordinately involved in the inhibitory effect on these essential steps of metastasis by MJ-29. Finally, our study provides evidence for G2/M phase arrest and apoptosis caused by HMJ-38 in CAL 27 cells, and HMJ-38 by intraperitoneal administration suppressed CAL 27 xenografts tumor growth in nude mice. Overall, we presented the novel findings that MJ-29 and HMJ-38 possess a potent therapeutic value for use against leukemia and oral cancer in response to cell-fate decision, and the efficacy of both compounds might be sufficient to investigate the potential of treatment in the future.
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