天然化合物誘導頭頸癌細胞產生細胞凋亡之機制探討

研究目的：根據統計資料，頭頸癌在全球與臺灣皆位於前十大癌症之列，好發於50-60歲之中壯年男性，除了影響個人健康，也會影響家庭經濟，甚而造成整體社會之疾病負擔。頭頸癌中以口腔癌最為常見，且其標準化發生率呈現逐年上升之趨勢；而鼻咽癌則是具有特殊之地理分布，好發於包含臺灣的東南亞與東亞一帶。因此本研究旨在尋找有潛力且低細胞毒性之天然化合物，探討其在對抗頭頸癌，特別是口腔癌與鼻咽癌方面之作用。研究方法及材料：本研究使用毛蘭素與薑花素D分別作用於人類鼻咽癌細胞株與口腔癌細胞株，並進一步分析其分子生物機制。進行細胞毒性試驗、細胞週期分析，利用DAPI染色法進行凋亡細胞之定量，也進行Annexin細胞染色法、粒線體膜電位分析，並利用西方墨點法測定蛋白質表現。研究使用GraphPad Prism Software Version 5.0 (GraphPad Software Inc., La Jolla, CA)，所有實驗至少重複3次，以ANOVA與Student's t test用於組別間之比較，P值＜0.05被認為具統計學上顯著意義。研究結果：研究結果顯示，毛蘭素具活化人類鼻咽癌NPC-NPC-039和NPC-BM細胞株凋亡與抑制細胞週期的作用，透過粒線體膜電位改變與死亡受體作用途徑，以毛蘭素處理過的鼻咽癌細胞中磷酸化ERK1 / 2呈劑量關係性減少，在ERK1 / 2抑制劑實驗中也可見相對應之結果，推測其為促進細胞凋亡的分子路徑；另一方面，薑花素D在人類口腔癌細胞株也具促進細胞凋亡之效果，實驗顯示其作用為透過JNK1 / 2之分子訊號途徑。結論與建議：本研究顯示，天然化合物毛蘭素與薑花素D分別在頭頸癌中的鼻咽癌與口腔癌細胞株具促進細胞凋亡之作用，屬於具發展潛力的天然抗癌製劑，未來有待進一步實驗確認其在動物與人體之成效。

Objective：According to global and Taiwan’s statistics, head and neck cancer is among the top ten cancers and more common middle-aged men, who are usually the main source of income for the family, so this creates a large burden of disease. Oral cancer is the most common among the head and neck cancers with a rising incidence rate; while nasopharyngeal cancer has a distinct ethnic difference and geographical distribution with higher inceidence rate Southeast Asia and East Asia including Taiwan. The purpose of this study is to determine whether the natural componds are effective against head and neck cancer cells and to establish their mechanism of action.Methods and Materials：In this study, Erianin and Coronarin D were used to act on human nasopharyngeal carcinoma cell lines and oral cancer cell lines, respectively. We perform cell cytotoxicity assay and cell cycle analysis. Annexin-V and PI double staining and DAPI staining were then done to evaluate the cell apoptosis level. Mitochondrial membrane potential measurement was used to confirm the mitochondria-mediated mechanism. Western Blot Analysis was also performed. Statistical analyses were performed using GraphPad Prism Software Version5.0 (GraphPad Software Inc., La Jolla, CA). ANOVA and Student's t test were used to compare the differences between groups and P values of<.05 were considered to represent statistically significant changes.Results：The result showed that erianin significantly increased activation of apoptosis in NPC-039 and NPC-BM cell lines and arrest the cell cycle obviously through mitochondrial membrane alternation, death receptors activation, and caspase-3, -8, -9 activation. Phosphorylated ERK1/2 was also decreased in erianin-treated NPC cell and the result was thought to promote apoptosis. Coronarin D also induces human oral cancer cell apoptosis though upregulate JNK1/2 signaling pathway. Conclusion and Suggestion：This is the first research to identify the anti-NPC effect of Erianin and anti-oral cancer effect of Coronarin D via the apoptosis mechanism. The natural compounds such as Erianin and Coronarin D were promising natural agents against head and neck cancer.