中藥植物石蟾蜍(Stephania tetrandra S. Moore)主成分漢防己甲素(tetrandrine)抑制人類口腔癌細胞離體和活體分子作用機轉

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题名:	中藥植物石蟾蜍(Stephania tetrandra S. Moore)主成分漢防己甲素(tetrandrine)抑制人類口腔癌細胞離體和活體分子作用機轉 Molecular Mechanism of Tetrandrine (Component from Chinese Herb Stephania Tetrandra S. Moore) on the Inhibition of Human Oral Cancer Cells in Vitro and in Vivo.
作者:	鍾景光(Jing-Gung Chung)
贡献者:	生命科學院生物科技學系;中國附醫醫學研究部
关键词:	口腔癌;漢防己甲素;細胞凋亡;細胞自噬;血管新生;oral cancer;tetrandrine;apoptosis;autophagy;angiogenesis
日期:	2013-07-31
上传时间:	2013-01-25 13:57:24 (UTC+8)
摘要:	目前口腔癌的治?方法以手術、放射線及化?為主，但化?藥物?效?人?甚滿意。近??眾多學者發現細胞自噬和癌症有很大的關?，因此該方向成為一種新興的抗癌治?方針。初步實驗結果發現，經漢防己甲素(tetrandrine)處?後的口腔癌細胞 (Cal27)，出現磷脂絲氨酸外翻、活化態的caspase-3、-8、-9 及-PARP 蛋白表現?上升，促使細胞凋亡的發生；同時間，也觀察到細胞內大?的酸性液囊泡產生，LC3II、Atg5、 Beclin-1 等自噬相關蛋白表現?增加，?明漢防己甲素會誘導細胞產生自噬的作用。這次計畫分為三大目標於三?內依序執?。第一?目標是?用漢防己甲素對人?口腔癌細胞誘導細胞凋亡與自噬作用的分子機轉探討。接著，?用正常小鼠找出漢防己甲素在體內?具毒性的有效劑?。第二?目標是確認漢防己甲素對?種移植口腔癌?鼠是否具有抗癌的活性。另外，也檢視漢防己甲素對arecoline 和 4-nitroquinoline 1-oxide 誘導口腔癌的?鼠是否有抗癌效果。第三?目標是探討漢防己甲素對人?臍帶靜脈內皮細胞和口腔癌細胞的血管新生之?體試驗。檢測漢防己甲素對人?臍帶靜脈內皮細胞和口腔癌細胞之存活?、血管內皮生長因子分?、血管新生相關蛋白表現的影響；並以小黑鼠體內血管新生的實驗檢測活體內血管新生的影響。最後，我們將提出漢防己甲素在活體和? 體試驗?影響血管新生可能的訊息傳遞?徑。 Currently, the treatment of oral cancer including surgery, radiotherapy and chemotherapy, but the chemotherapy is still unsatisfied. Numerous evidences already shown that autophagy has associated with the cancer development. Thus, agent to induce autophagy is the trend for anticancer. Primary results indicate the tetrandrine induced apoptosis of Cal27 human cancer cells via increase of phosphotidylserine exposed on the surface of the cell; promote caspase-8, -9 and -3 activities, and increased p-PARP expression. At the same time, results also showed acidic vacuoles present, increase the expression of LC3II, Atg5 and Beclin-1 that associated with autophagy. Based on these primary observations, we suggest that tetrandrine may induce cell death through the induction of autophagy in Cal27 cells. This proposal is divided into three subjects which will be done in three years. Subject I in the first year is to investigate the effects of tetrandrine on Cal27 human oral cancer cells, then to find out the molecular signal pathways of apoptosis and/or autophagy. Finally, to investigate the cytotoxitic effect on the normal animal model in vivo. Subject II in the second year to examine the efficiency of tetrandrine on the anticancer activity in vivo and it can be used nu/nu mice for examining Cal27 cells into xenograft nu/nu BALB/c mice. Furthermore, we will use chemical carcinogens such as arecoline and 4-nitroquinoline 1-oxide to generate oral cancer mice, then to examine the death of oral cancer by tetrandrine. Subject III in the third year is to investigate the effect of treatment on angiogenesis from HUVECs and human liver cancer cells in vitro. We will examine the agent affecting the total viable cells, secretion of VEGF, angiogenesis associated genes’ expression and associated protein levels. Furthermore, We will examine those agents affecting angiogenesis in vivo based on Matrigel plug assay. Finally, we examine the tetrandrine effect on angiogenesis associated genes' expression and proteins, then to find out the possible signal pathway in vitro and in vivo.
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