大黃酚誘發人類肝癌細胞株(SK-HEP-1與J5)細胞壞死透過活性氧化物質與鈣離子產生和ATP層次的影響; Chrysophanol induced necrosis through the productions of reactive oxygen species, calcium and affects ATP levels in human liver cancer SK-HEP-1 and J5 cells

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题名:	大黃酚誘發人類肝癌細胞株(SK-HEP-1與J5)細胞壞死透過活性氧化物質與鈣離子產生和ATP層次的影響;Chrysophanol induced necrosis through the productions of reactive oxygen species, calcium and affects ATP levels in human liver cancer SK-HEP-1 and J5 cells
作者:	呂啟誠;Chi-Cheng Lu
贡献者:	中國醫藥大學：醫學研究所碩士班
关键词:	大黃酚;人類肝癌細胞(SK-HEP-1與J5);細胞壞死;三磷酸腺苷;Chrysophanol;Human liver cancer cells (SK-HEP-1 & J5);Necrosis;ATP;Reactive oxygen species
日期:	2007-06-22
上传时间:	2009-08-13 14:50:50 (UTC+8)
摘要:	肝癌是世界上最普遍的惡性腫瘤之一，尤其是在台灣。因此，抗癌藥物能夠引起肝臟最小傷害或延滯至肝硬化是有效認為治療肝癌決定性的方法。大黃酚Chrysophanol （1, 8-dihydroxy-3-methyl -anthraquinone）是一種衍生來自已被使用久遠的傳統中藥「大黃」的蔥??類（anthraquinone）化合物。到目前為止，尚無有用的大黃酚影響肝癌細胞株（SK-HEP-1與 J5）的文獻資料或報告可供參閱。本實驗研究中，我們利用實驗說明大黃酚誘發人類肝癌細胞株均有產生毒殺的影響，而且利用流式細胞技術分析顯示，並無明顯產生細胞凋亡的現象。然而，兩株人類肝癌細胞均有活性氧化物質（ROS）與胞內鈣離子增加的現象，且粒線體膜電位（ΔΨm）呈現下降的結果。人類肝癌細胞（SK-HEP-1與 J5）在暴露於大黃酚後，ATP層次會有下降的效果。亦利用西方墨點法與反轉錄聚合?○s鎖反應用來檢測有關本實驗的蛋白質與基因層次的表現；也使用General caspase抑制劑（z-VAD-fmk）、ROS抑制劑（NAC）與細胞壞死（Necrosis）抑制劑（IM-54）和鈣離子螯合劑（BAPTA）來確認我們的實驗最後結果。經由實驗數據發現，大黃酚能誘發人類肝癌細胞株（SK-HEP-1與 J5）產生細胞壞死（Necrosis）的現象出現。 Hepatocellular carcinoma (HCC) is one of the most common and malignancies in the world, especially in Taiwan. Therefore, the anti-cancer agents that cause minimal damage to cirrhotic liver are considered crucial for successful treatment of HCC. Chrysophanol (1, 8-dihydroxy-3-methyl-anthraquinone) is one of the anthraquinone derivatives from rhubarb (Da huang) which had been used a traditional Chinese medicine since long ago. There is no available information to address the effect of chrysophanol on human liver cancer SK-HEP-1 and J5 cells. In this study, we demonstrated that chrysophanol induced cytotoxicity in examined both cell lines. Flow cytometric analysis did not show apoptosis (sub-G1 group). However, the levels of reactive oxygen species (ROS) and intracellular Ca2+ were increased, but the level of mitochondrial membrane potential (ΔΨm) is decrease in examined both hepatoma cell lines. ATP levels were decreased in both examined cells after exposed to chrysophanol. Western blotting and RT-PCR were used to detect the associated protein and gene levels. We also used general caspase inhibitor (z-VAD-fmk), ROS inhibitor (NAC), necrosis inhibitor (IM-54) and Ca2+ chelator (BAPTA) to confirm our results. Based on those findings indicated that chrysophanol induced necrosis in both examined human liver cancer cells.
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