利用cDNA微陣列整合平台研究樟芝菌絲體萃取物對KG-1 細胞株基因調控之影響

The Study of Gene Expression Profiles of KG-1 Cells Treated with Antrodia Camphorata Mycelia Extracts using cDNA microarray

彭維義(Wai-Yi Pong)

指導教授 ： 黎耀基 ； 許志[]

國立清華大學/生命科學院/生物科技研究所/碩士(2006年)

摘要

樟芝是一種用來抗癌和抗發炎的台灣民間著名醫藥蕈類 (無褶菌目，多孔菌科)。然而，目前對於不同樟芝萃取物影響白血病細胞生長抑制的分子機制尚未明瞭。在本研究中，我們結合草藥鑑定、UV/VIS光譜法、高效液相層析法與感應偶合電漿質譜法建構一個整合性高效率平台，來將草藥藥材進行標準化。同時，我們以cDNA微陣列科技來檢視不同樟芝萃取物(水萃與醇萃物)對KG-1(急性骨髓性白血病細胞株)細胞的生長抑制影響。基於此科學平台，我們發展出一個可行的定量方法並建立出有機/無機的化學指紋。透過比較草藥萃取物對KG-1整體基因的表現型式來看，數個可能的生物反應與相關的標的基因，將可被以基因本體論(Gene Ontology)為基礎的群集方法所鑑定出來。從兩種萃取物的基因本體論歸類比較之顯著性來看，結果暗示樟芝水萃物會下調分解與碳水化合物代謝相關的基因群，而樟芝醇萃物則會促進MHC class II 抗原呈現相關基因群的表現。

關鍵字

樟芝 ； 菌絲體 ； 萃取物 ； cDNA微陣列 ； 分子標的 ； 平台

並列摘要

Antrodia camphorata ( A. camphorata ) is well known in Taiwan as a folk medicinal fungus with anticancer and anti-inflammatory effects. However, the molecular mechanism underlying leukemia growth inhibition effects through different A. camphorata treatment is poorly understood. In this study, we constructed an integrated high-throughput platform to standardize the herbal materials by using a combination of herbal identification, UV-VIS spectrophotometry, high performance liquid chromatography (HPLC), and inductively coupled plasma mass spectrometry (ICP-MS). Simultaneously, we used cDNA microarray technology to examine the effects of various A. camphorata mycelia extracts (water and methanol extracts) on the effects of growth inhibition on KG-1 cells (human acute myelogenous leukemia cell line). Based on this science-based platform, we developed a practical quantitative method and established an organic/inorganic chemical fingerprint. By the comparison of KG-1 global gene expression patterns from herbal extracts, several possible biological responses and relevant marker genes were identified by gene ontology (GO)-based clustering analysis. The statistically significantly comparison of GO category from both treatments indicated that water extract could down-regulate carbohydrate catabolism related genes and methanol extract up-regulate MHC class II antigen processing genes, respectively.

參考文獻

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