阿魏酸與其衍生物對於人類非小細胞肺癌之生長與轉移能力機制探討

The effect of ferulic acid and its derivative on proliferation and cellular migration of human non-small cell lung cancer

胡惠婷(Huei-Ting Hu)

指導教授 ： 邱建智

高雄醫學大學/生命科學院/生物科技學系碩士班/碩士(2012年)

https://doi.org/10.6832/KMU.2012.00119

摘要

肺癌是目前位居國人癌症死亡原因之首，而非小細胞肺癌 (non-small cell lung cancer, NSCLC) 占肺癌中達80%之多，其中的大細胞肺癌 (large cell carcinoma) 具生長快速、高度轉移能力及多重抗藥性的特性，病患經治療後有預後情形較差的現象，因此找出新穎藥物具有效對抗大細胞肺癌是極為重要的目標。酚類化合物 (phenolic compounds) 廣泛分佈於蔬菜和水果之中，其主要的芳香環及羥基結構經證實是具有抗氧化效力的指標。近年來酚類化合物經研究證實具有治療人類常見疾病的潛力，包括治療神經退行性疾病、癌症、糖尿病、心血管功能障礙、發炎症和老化。阿魏酸 (ferulic acid, FA) 就屬於天然酚類化合物，經研究證實具有抗氧化的能力。然而，目前阿魏酸的研究大多著重在抗氧化層面的探討，但對於癌症的細胞生理機制所知極為有限。我們的研究目的是要探討阿魏酸對人類肺癌H1299細胞之生理影響。在試管內 (In vitro) 抗氧化測試顯示，FA對於DPPH自由基清除劑表現出顯著抗氧化作用。然而本實驗顯示FA處理能增加H1299肺癌細胞內生性活性氧，包含過氧化氫 (H2O2) 和超氧陰離子自由基 (O2-) 。即使FA不誘導H1299細胞凋亡，但在細胞群落形成分析 (colony formation) 與細胞非依賴性貼附 (anchorage-independent growth) 實驗顯示，FA能顯著抑制細胞增殖，這表示FA可能具有長期對抗肺癌的潛力。此外，FA會導致促進增生的轉錄因子β-catenin在胺基酸Ser33, Ser37, Thr41和Ser45磷酸化，使得磷酸化的β-catenin被蛋白酶體分解。另外，FA降低MMP-2/-9活性，進而影響H1299移動與侵襲的轉移能力，因此綜合實驗結果得知FA對於肺癌可能具有化學預防 (chemoprevention) 的潛力。

最近，從蔬菜茭白筍殼 (Zizania latifolia coba husk) 分離出一個阿魏酸衍生物- Feruloy-L-arabinose (FAA)。結果顯示，FAA在抗氧化能力程度較FA佳，且比FA更具親水性 (hydrophility)，FA短期處理對H1299細胞並無毒性。此外，細胞群落形成與細胞非依賴性貼附生長試驗也顯示，FAA與FA同樣能夠抑制細胞增殖的情形。細胞內生性活性氧含量顯示，FAA皆強烈清除細胞內生性活性氧H2O2與O2-之含量，且對細胞移動和侵襲的抑制作用效果比FA佳。總而言之，我們目前的工作提供了一個阿魏酸衍生物在肺癌細胞的細胞移動之新功能，這表明FAA是未來發展有望的化學預防作用。

關鍵字

阿魏酸 ； 阿魏酸衍生物 ； 自由基 ； 活性氧 ； 肺癌 ； 細胞轉移 ； 化學預防

並列摘要

Phenolic compounds are widely distributed in the plants, especially in vegetables and fruits. Phenolic compounds are considered as potential therapeutic agents against a variety of human diseases, including neurodegenerative diseases, cancer, diabetes, cardiovascular dysfunction, inflammatory diseases and ageing. Ferulic acid (FA), a naturally phenolic compound, was reported to exert antioxidant effects in vitro. However, the mechanism of FA on cellular physiology, especially the cancer physiology, remains largely unknown. The aim of our study is to investigate the cellular physiological effects of FA and its derivative on human lung cancer cells. The DPPH free radical scavenging experiments showed the potent antioxidant effect of FA in vitro. But FA increased the endogenous reactive oxygen species (ROS), namely H2O2 and O2-, in lung cancer H1299 cells. Despite FA did not affect the cell viability and induce the apoptosis, the colony formation assay presented that FA significantly attenuates the cellular proliferation, suggesting the long-term anti-cancer effect of FA. Furthermore, FA inhibited the migration of lung cancer cells. Interestingly, the inhibitory effect of FA was observed at the doses of 0.15, 0.3 and 0.6 mM but not at lower dose, indicating that the anti-cancer effect of FA is dose-dependent. In addition to, the Western blotting results showed that the FA caused the phosphorylation of β-catenin (on residues Ser33, Ser37, Thr41 and Ser45), a critical transcription factor of cellular migration and proliferation. The phosphorylation results in the proteasomal degradation of β-catenin. Recently, a new FA derivative, feruloyl-L-arabinose (FAA) was isolated from the vegetable coba husk. Our results showed the FAA was more hydrophilic than FA. Like FA, FAA exerts no significant cytotoxicity to H1299 cells. In comparison with FA, FAA exerts a stronger scavenging effect of intracellular ROS and inhibitory effect on cellular invasion and migration. Taken together, our present work provided a new insight into the novel function of FAA on cell migration in lung cancer cells, suggesting the promising role of the FAA in chemoprevention.

並列關鍵字

ferulic acid ； feruloyl-L-arabinose ； free radical ； reactive oxygen species (ROS) ； lung cancer ； migration ； chemoprevention

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