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研究生(外文):Min-Hsuan Lin
論文名稱(外文):The anti-cancer efficacy of pyrimethamine on non-small cell lung cancer
指導教授(外文):CHI-CHEN LIN
外文關鍵詞:Non small cell lung cancersPyrimethamineapoptosisautophagy
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目前已有許多腫瘤的研究指出,乙胺嘧啶(PYR)不論在體內或體外的試驗都具有抗腫瘤的效果,但唯獨沒有關於非小細胞肺癌(NSCLC)的研究。因此在本研究中,我們證明了PYR可以導致非小細胞肺癌的細胞株A549細胞存活率下降,並且可誘導A549細胞的G1期細胞產生停滯,以及大量細胞在sub-G1期的堆積。利用Annexin V和PI雙重染色顯示,在PYR作用下的A549細胞會發生凋亡的情形,證實PYR除了可以誘導細胞在sub-G1期堆積外,此外PYR還可以使A549產生pro-survival細胞自噬。綜合我們的研究結果顯示,PYR可以誘導細胞凋亡,同時也將誘導細胞產生自噬的狀況,未來可將PYR用來化療或治療非小細胞肺癌。

Despite several studies had reported the anti-tumor effect of Pyrimethamine (PYR) in vitro and in vivo in many tumors, there is none about non-small cell lung cancer (NSCLC). In this study, we demonstrated that PYR reduces the cell viability of NSCLC cell lines A549 and induces G1 phase arrest in cells. Double staining of Annexin V and PI indicated that PYR could induce apoptosis in A549 cell, PYR induces sub-G1 phase accumulation. Furthermore, PYR can also induce the pro-survival autophagy in A549 cells. These results suggested PYR induces G1 phase arrest apoptosis to affect the proliferation of A549 cells. PYR also induces the pro-survival autophagy to maintain A549 survival. In summary, our findings showed that PYR promotes intrinsic apoptosis by inducing apoptotic protein activation and can also induces autophagy, suggesting PYR may be considered as chemo-prevention or treatment NSCLC.

摘要 i
Abstract ii
目次 iii
圖表目次 v
縮寫表 v
第一章 緒論 1
一、 肺癌(Lung cancer) 1
二、 乙胺嘧啶之簡介(an introduction of Pyrimethamine) 3
三、 細胞週期(Cell cycle)3
四、 細胞凋亡(Apoptosis) 3
五、 細胞自噬(Autophagy) 6
第二章 研究動機 7
第三章 實驗材料與方法 8
一、 方法步驟 8
1. 細胞繼代培養(subculture of cells)8
2. 計算細胞數 8
二、 細胞存活率檢測(MTT assay) 9
三、 細胞凋亡分析(Annexin V/PI staining assay) 10
四、 細胞週期分析(Cell Cycle assay)10
五、 酸性囊泡器檢測(Acidic vesicular organelles detection) 11
六、 分析與統計方法(Statistical analyses)11
第四章 結果 12
一、 Pyrimethamine (PYR)能有效抑制A549人類肺癌細胞生長作用 12
二、 Pyrimethamine (PYR) 對A549人類肺癌細胞生長抑制走向細胞凋亡主導之型式 12
三、 Pyrimethamine (PYR) 對A549人類肺癌細胞之細胞週期影響 12
四、 Pyrimethamine (PYR) 對A549人類肺癌細胞之酸性囊泡器檢測 12
第五章 研究討論 14
第六章 參考文獻 16
第七章 實驗結果圖 19
圖一、Pyrimethamine對於非小細胞肺癌A549細胞之增生作用影響 19
圖二、Pyrimethamine對於非小細胞肺癌A549之細胞凋亡的影響 20
圖三、Pyrimethamine對於非小細胞肺癌A549之細胞週期分佈 21
圖四、Pyrimethamine對於非小細胞肺癌A549之酸性囊泡器檢測 22

附圖 1主要癌症死亡原因 2
附圖 2半胱氨酸蛋白酶(Procaspases)活化途徑 2
附圖 3凋亡途徑與內源性凋亡途徑 5
附圖 4凋亡蛋白酶活化途徑 5

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