大黃素、蘆薈大黃素、大黃酸對人類舌鱗狀細胞癌細胞株（SCC4）的細胞週期停滯及細胞凋亡之影響; Effects of emodin, aloe-emodin and rhein on cell cycle arrest and apoptosis in human tongue squamous carcinoma cell line (SCC4)

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题名:	大黃素、蘆薈大黃素、大黃酸對人類舌鱗狀細胞癌細胞株（SCC4）的細胞週期停滯及細胞凋亡之影響;Effects of emodin, aloe-emodin and rhein on cell cycle arrest and apoptosis in human tongue squamous carcinoma cell line (SCC4)
作者:	賴婉文;Wan-Wen Lai
贡献者:	中國醫藥大學：醫學研究所碩士班
关键词:	大黃素;蘆薈大黃素;大黃酸;人類舌鱗狀細胞癌;細胞凋亡;emodin;aloe-emodin;rhein;human tongue squamous carcinoma;apoptosis
日期:	2007-06-20
上传时间:	2009-08-13 14:50:26 (UTC+8)
摘要:	中文摘要口腔癌是一種發生在口腔中的一種惡性腫瘤。在美國疾病管制及預防中心的統計中顯示每年都會新增30000個口腔癌新案例。而在台灣的統計調查，發現口腔癌已躍居十大癌症的第四名。鱗狀癌是最常見的惡性腫瘤號發於腸胃道以上，而它佔口腔癌的比例達90-95％。大黃是ㄧ種傳統中藥，已早在很久以前被沿用於各種中藥調製至今。大黃素、蘆薈大黃素及大黃酸是在大黃中最重要的成分，皆為蒽醌類化合物。這三種化合物已被證實具有數種的生物活性作用，例如:抗病毒、抗菌、刺激細胞激素的釋放、保肝及對於人類癌細胞有抗腫瘤的活性等。但目前這三種化合物對於人類舌鱗狀細胞癌(SCC4)的研究探討尚未有相關的資訊。因此本篇研究我們想探討大黃素、蘆薈大黃素及大黃酸對於人類舌鱗狀細胞癌(SCC4)做相關的體外試驗來探討是否有抗癌作用的能力。我們的研究結果顯示大黃素、蘆薈大黃素及大黃酸在特定的時間處理人類舌鱗狀細胞癌(SCC4)，其細胞存活率下降。並這些化合物會誘導SCC4細胞的細胞週期停滯（大黃素：G2-M期、蘆薈大黃素:S期、大黃酸:S期）及細胞凋亡。其發現利用流式細胞儀檢測發現，活性氧化物（ROS）產生及鈣離子釋放，並促進Caspase-3,-8,-9活性產生並降低粒線體膜電位（MMP）。另外也利用DNA fragmentation、DAPI染色、confocal觀察等實驗，其結果皆有細胞凋亡的象徵。並利用西方墨點法來檢測其細胞的週期蛋白及凋亡蛋白表現之路經機轉。發現大黃素會促進粒線體路徑及ER stress路徑走向細胞凋亡；蘆薈大黃素會促進粒線體路徑、死亡受體路徑及ER stress路徑走向細胞凋亡；大黃酸促進死亡受體路徑走向細胞凋亡。從以上的實驗結果發現大黃素、蘆薈大黃素及大黃酸對於人類舌鱗狀細胞癌(SCC4)具有誘導細胞凋亡的能力，因此推測這三種化合物對人類舌鱗狀細胞癌細胞(SCC4)可能具有抗癌的作用。 The oral cancer is an occurrence in the malignant tumor of the oral part. The Centers for Disease Control and Prevention say there are 30,000 new cases of oral cancer each year in USA. The oral cancer has already reached the fourth greatest cancers causes death in Taiwan. Squamous carcinoma is the most familiar malignant tumor in upper gastrointestinal tract. It account for 90%-95% of oral cancer. Rhubarb is a traditional Chinese medicine which has been used since ancient times and is still present today in various herbal preparations. Emodin, aloe-emodin and rhein are major components of rhubarb. The three compounds have been demonstrated that possess a number of biological activities such as anti-virus, anti-bacteria, stimulation of cytokine release, vasorelaxative, hepatoprotective, and anti-tumor activity in human cancer cell lines. There is no available information to address those three compounds effects on human tongue squamous carcinoma cell line (SCC-4). Therefore, in this study, we want to examine whether or not emodin, aloe-emodin and rhein have an anti-cancer ability of repressing the SCC4 cancer cells in Vitro. The results demonstrated that three examined compounds induced the cell cycle arrest (emodin: G2-M phase arrest; aloe-emodin and rhein: S phase arrest) and apoptosis in SCC-4 cell line. Three compounds also increased the reactive oxygen species and Ca2+ production, promoted Caspase-3, -8 and -9 activities and decreased the levels of mitochondrial membrane potential in examined SCC-4 cells. Western blotting analysis revealed that aloe-emodin promoted p53, p21 and p27 and inhibited the levels of Cyclin E and CDK2 before leading to S phase arrest. Those effects are dose time-dependent manners. Aloe-emodin promoted Caspase-3 and -9 activities, increased the levels of Bax and decreased the levels of Bcl-2. It also promoted Caspase-8 activity increased the levels of FasLigand. The western analysis maybe demonstrated that course to increase the production ROS and decreasing the levels of mitochondrial membrane potential before being induced apoptosis.
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