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研究生:張兆嶔
研究生(外文):Chao-Chin Chang
論文名稱:蘆薈大黃素對舌咽癌細胞株之凋亡和侵犯機制的影響
論文名稱(外文):The Effects of Aloe-Emodin on Apoptosis and Invasion of the Pharyngeal Squamous Carcinoma Cell Line
指導教授:蘇鴻麟蘇鴻麟引用關係
指導教授(外文):Hong-Lin Su
學位類別:碩士
校院名稱:國立中興大學
系所名稱:生命科學系所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:68
中文關鍵詞:蘆薈大黃素舌咽癌細胞
外文關鍵詞:aloe emodin
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雖然有證據證明蘆薈大黃素具有成為抗癌藥物的潛力,但蘆薈大黃素引起人類舌咽癌細胞株凋亡的機制卻尚未瞭解。在本篇研究中首先證實了蘆薈大黃素能夠抑制舌咽癌細胞株的增生並且誘發其凋亡,在蘆薈大黃素所引發的細胞凋亡中可以清楚的觀察到細胞核的濃縮與染色體的裂解,蘆薈大黃素所引起的細胞凋亡伴隨著Bax表現量的上升和Bcl-XL表現量的下降,而Bcl-2的表現量則無顯著變化。在以蘆薈大黃素處理過的舌咽癌細胞中還觀察到caspase-3活性的提高,粒線體膜電位下降,造成cytochrome c和AIF自粒線體轉位至細胞質,並且提高了細胞質中ROS和鈣離子的含量。這些結果說明了蘆薈大黃素在舌咽癌細胞中經由粒線體所調控的途徑誘發舌咽癌細胞進行細胞凋亡;另外,它還能藉由降低MAPK訊息傳導路徑中上游重要的蛋白質的表現量,如Grb2和SOS1,使得下游的轉錄因子調控的基因,如MMP-9,的表現量下降,進一步造成癌細胞移轉的能力下降。以上結果說明了蘆薈大黃素可能作為一種有效治療舌咽癌細胞的抗癌藥物。
Although there are evidences showing that aloe-emodin might be a potential anticancer drug, but the mechanisms that involved in its action on the induction of apoptosis in human pharyngeal squamous carcinoma(PSC)cells are remained to defined. In the present study, we first demonstrated that aloe-emodin could inhibit proliferation and induce apoptosis in PSC cells. Aloe-emodin-induced apoptosis was clearly verified by the appearance of nuclear condensation and DNA fragmentation. Apoptosis induced by aloe-emodin was accompanied by the up-regulation of Bax and down-regulation of Bcl-XL, but no effect on the level of Bcl-2. A significant increase in caspase-3 activity was observed in aloe-emodin treated PSC cells. Moreover, we found that aloe-emodin induced the loss of mitochondrial membrane potential(MMP), the release of cytochrome c and apoptosis-inducing factor(AIF)from mitochondria to cytosol. And also resulted in an elevation of reactive oxygen species (ROS) and increase calcium (Ca++) level in cytosol. In addition to inducing apoptosis, aloe-emodin inhibited the expressions of important proteins in Ras-MAPK signaling pathway, Grb2 and SOS1, and the genes regulated by the signaling pathway were suppressed. As the genes regulated by Ras-MAPK signaling pathway, MMP-9 and VEGF, were down-regulated, the invasion activity of cancer cells was suppressed. These results suggest that aloe-emodin induced apoptosis and inhibited invasion in PSC cells were mediated through mitochondria dependent and Ras-MAPK pathways. It implies the possibility that
aloe-emodin could be a useful anticancer agent for treatment of PSC.
總目錄
摘要……………………………………………………………………Ⅰ
Abstract………………………………………………….………….Ⅱ
第一章 前言……………………………………………………………1
第二章 文獻探討………………………………………………………3
第一節 癌症……………………………………………………………3
第二節 簡介蘆薈大黃素(aloe emodin)……………………….…4
第三節 細胞凋亡(Apoptosis)……………………………………6
第四節、細胞遷移(Cell Migration) ………………………………8
第三章 材料與方法……………………………………………….10
第一節 實驗材料…………………………………………………10
第二節 實驗方法………………………………………………14
第四章 結果…………………………………………………………… 30
第五章 討論……………………………………………….…….36
第六章 結論……………………………………………………………40
第七章 附錄……………………………………………………………62
第八章 參考文獻………………………………………………………65

圖目錄
圖1. 蘆薈大黃素對舌咽癌細胞的毒性………………………………42
圖2. 蘆薈大黃素造成舌咽癌細胞之細胞形態的變化………………43
圖3. 加入蘆薈大黃素之後(A)以DAPI染舌咽癌細胞的細胞核並觀察其形態變化 (B)以單細胞凝膠電泳觀察細胞核中DNA裂解程度的影響………………………………………………………………....44
圖4.觀察加入蘆薈大黃素之後(A)對舌咽癌細胞內caspase 3蛋白活性的影響 (B)對pro-caspase-3裂解程度的影響……………………….45
圖5.觀察加入蘆薈大黃素之後對舌咽癌細胞內p53蛋白的活化(A)蘆薈大黃素對Bcl-XL 和p53的表現量的影響(B and C)p53和p21轉錄活性的活化………………………………………………..….46
圖6. 以流式細胞儀觀察加入蘆薈大黃素之後對舌咽癌細胞內(A)粒線體膜電位(MMP)(B)活性氧化物含量(C)鈣離子濃度的影響………………………………………………………………..…47
圖7. 觀察加入蘆薈大黃素之後粒線體釋放(A)cytochrome c和(B)AIF的程度…………………………………………………..……49
圖8. 蘆薈大黃素對舌咽癌細胞侵犯能力的影響…………………….52
圖9. 蘆薈大黃素抑制舌咽癌細胞遷移的能力…………………….54
圖10. 蘆薈大黃素能降低舌咽癌細胞的癒傷能力……………………57
圖11. 蘆薈大黃素對MMP-9的蛋白表現量的影響……………………58
圖12. Grb2/SOS-Ras-MAPK所調控的訊息傳導受到蘆薈大黃素的抑制…………………………………………………………………..…60
圖14. 處理40 μM蘆薈大黃素之後NF-κB在細胞核及細胞質中含量的變化………………………………………………………………61
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