桑黃輔助癌症化學治療與評估改善多重藥物抗藥性表現之藥理機制探討
褚俊傑
南台科技大學
桑黃的學名為Phellinus linteus，是寄生於桑樹上的一種多年生蕈類，生長緩慢，難以用人工方法栽培,在韓國與日本已被採用作為醫治癌症的藥物。近年研究發現，桑黃具有抗腫瘤及免疫調節等功效。在最近這幾年中，人們開始注意到桑黃 (Phellinus Linteus) 這種藥用蕈對醫治癌症的特效性。許多日本及韓國的醫學研究者早已經確認桑黃具有絕佳的抗癌力（抗腫瘤活性）。於一九八○年代時韓國跟著最先研究桑黃的日本開始對野生桑黃進行正式的研究。直至一九九三年，韓國政府終於正式許可桑黃成為第一種以蕈類作為抗癌劑的醫藥品。及至數年前，日本也將桑黃當成健康食品，正式進口販售，於是使用桑黃者大大激增。由於桑黃的免疫與抗腫瘤功效且不具有副作用的優點，不過因桑黃生長緩慢所以數量有限，而本校利用發酵的技術將桑黃不再是昂貴及稀少的抗癌劑，不過必須經過多方實驗印證後才能對本校所生產的桑黃發酵菌絲體具有相同的抗癌效果。 因此，本計劃擬研究桑黃發酵菌絲體與抗癌藥物Cisplatin, Etoposide, Methotrexate, Mitomycin及Paclitaxel相互評估，投予黑色素細胞癌、肺癌與肝癌等腫瘤動物模式：腫瘤接種模式、誘導原發性肝癌動物模式及靜脈注射誘導腫瘤模式，測試桑黃是否能輔助抗癌藥物更可以有明顯的抗癌作用。首先，由誘導癌症實驗動物透過抽取血液，酵素免疫沉澱分析法的方式來測量癌胚抗原(CEA)、甲型胎兒蛋白(AFP)、一氧化氮(NO)、自由基(superoxide, H2O2)與癌症相關之cytokines的濃度如TNF-α、IL-12和IFN-γ等的變化。之後利用流式細胞儀的方式來測量癌症細胞程式性凋亡的情形，再以免疫組織染色 ( immuno- histochemistry staining , IHC)、RT- PCR及西方墨點法來偵測相關指標性產物的量如NF-κB、PARP-1、AKT與MDR-1的表現。最後，進行實驗動物之存活率、腫瘤大小與腫瘤癌化指標(tumor biomarker)之觀察，透過腫瘤增生指標(BrdU incorporation)和細胞程式性死亡(TUNEL assay)等免疫組織染色檢查，來評估桑黃是否具有輔助其他相關抗癌藥物較佳的療效並探討桑黃之抗癌藥理活性與分子機制。
關鍵字：桑黃；黑色素細胞癌；肺癌；肝癌；腫瘤動物模式
The molecular mechanism of Phellinus linteus play an adjuvant role on a variety of anti-cancer drugs and improve multidrug resistance expression in animal models
Jiunn-Jye Chuu
Southern Taiwan University
Phellinus linteus, a unique fungal species, used as a folk medicine for over one thousand years. Hepatocellular carcinoma and lung cancer are the most prevalent cancers in Taiwan, often respond with limited efficacy to currently available chemotherapeutics with multidrug resistance (MDR) expression. The present study were to establish a valuable system in vitro and in vivo, compare the adjuvant anti-cancer effects from Phellinus linteus mycelia on the human hepatoma cell line (HepG2), mouse lung cancer cell line (LL2) and melanoma cell (B16F10). First, the cancer cells occurred resistance to several clinical chemotherapies, then the adjuvant anti-cancer effects of Phellinus linteus were observed by MTT assay and flow cytometry in vitro assay . Using 5-bromo-2’-deoxyuridine incorporation (for proliferation detection) and Annexin V-FITC/PI staining technology (for apoptosis detection), through modulating the MDR phenotype, which cause resistance to several malignant/recurrent tumors before surgical operation to evaluate the anti-cancer effects of Phellinus linteus extract. An in vivo-like three-dimensional culture (histoculture system) will been applied to evaluate the activity of anticancer drugs and its results are correlated to the clinical outcome in several solid carcinomas.
關鍵字：Phellinus linteus；melanoma；lung cancer；hepatoma
