紅麴菌 Monascus purpur CWT715 發酵液之抗轉移及抗氧化作用

In vitro effects of Monascus purpureus CWT715 on anti-metastasis and antioxidantion activity.

李婉如(Wan-Ju Lee)

指導教授 ： 黃晉修 ； 張駿志

亞洲大學/健康學院/保健營養生技學系碩士班/碩士(2010年)

摘要

癌細胞的轉移擴散是癌症病人致死的主要原因。研究證實，氧化壓力是造成癌症的原因之一，而抗氧化劑則具有化學預防 (chemoprevention) 的功效。紅麴有很好抗氧化能力，尤其二次產物更具許多保健功效，例如紅麴色素的抗癌作用以及dimerumic acid 的抗氧化能力等。研究證實紅麴具有抑制癌細胞生長的能力，但對於抑制轉移是否具有功效並不清楚，因此本研究探討紅麴菌 (Monascus purpureus CWT715) 發酵液的抗癌轉移作用，並釐清其作用機制。首先以酒糟作為營養源生產紅麴的二級代謝產物作為本研究樣品來源，並分別以人類肝癌細胞株 (SK-Hep-1)及小鼠胚胎肝臟細胞 (BNL CL.2) 作為研究模式，分別進行抗癌與抗轉移試驗以及抗氧化力試驗。並進行以下分析：細胞存活率、細胞侵入、細胞移行和基質金屬蛋白水解酵素 (matrix metalloproteinases, MMPs) 的活性及彗星影像分析(Comet assay)。結果發現：紅麴發酵液 (50 g/mL; 1~7天) 可顯著抑制SK-Hep-1肝癌細胞生長，其中以發酵第六天的紅麴液最具效果，在與肝癌細胞共同培養24小時後，其抑制率達到60% (p＜0.05)。另外，第六天紅麴發酵液也具有較高的色素和總酚含量，因此本試驗以第六天發酵液進行往後試驗。在培養不同劑量 (1~100 g/mL) 紅麴發酵液24和48小時後，在抑制癌細胞生長的作用上與劑
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量和時間呈正相關，在抗癌細胞轉移部份，紅麴發酵液可顯著抑制癌細胞侵入和移行，且呈劑量關係，在100 g/mL時可抑制侵入達68% (p＜0.05)，以及抑制移行達46% (p＜0.05)。由於紅麴對胞外酵素MMP-2和MMP-9的活性並無顯著抑制作用，因此推測紅麴抗轉移作用主要以抑制癌細胞移行有關。在抗氧化研究部分：預培養紅麴後再以0.3mM tert-butyl hydroperoxide (t-BHP)培養1小時誘導正常肝細胞 (BNL CL.2)氧化傷害，發現第七天紅麴發酵液可降低t-BHP 所造成之DNA損傷，且呈劑量關係，在劑量100 g/mL時可降低約48%。本研究證明，紅麴發酵液藉由抑制癌細胞移行進而達到抗轉移的作用，且對正常肝細胞有抗氧化之功效。

關鍵字

紅麴菌 ； 抗氧化 ； 抗癌 ； 抗癌轉移

並列摘要

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide and one of the leading causes of cancer death in Taiwan. Tumor metastasis is a major cause of death for cancer patients. It is well understood that oxidative stress is intricately associated to hepatic carcinogenesis. Although Monascus pigment has been shown to inhibit cell growth of Breast cancer, the anti-metastatic action is poorly understood. In the present study, we have focused on the anti-metastatic effects and mechanisms of the fermentation of M.purpureus CWT75. Here, we used SK-Hep-1 cells (a highly invasive hepatoma cell line) and BNL.CL 2 (mouse liver cell line and be used as normal liver cell) to test whether the fermentation of M.purpureus CWT75 has anti-cancer, anti-metastasis and antioxidants activity. After incubation with different fermentation of M.purpureus CWT75 (1-100 g/mL) for 24 and 48 h, we determined migration, invasion, and matrix metalloproteinases (MMPs) activities in SK-Hep-1 cells, and examined lipid peroxidation and DNA strand breakage (measured as comet assay) in tert-butyl hydroperoxide (t-BHP)-induced oxidative damage model in BNL.CL 2 cells. We showed that incubation of cells with the fermentation of M.purpureus CWT75 (1-100 g/mL) for 24 and 48 h resulted in significant inhibition of invasion and migration in SK-Hep-1 cells and that these effects were concentration-dependent. At 100 g/mL, the fermentation of M.purpureus CWT75 significantly inhibited cell migration and cell invasion by 46 and 68%, respectively. In addition, the fermentation of M.purpureus CWT75 also inhibited lipid peroxidation (thiobarbituric acid reactive substances, TBARS) and DNA stand breaks, with 51% (P<0.05) and 61% (P<0.05) suppression, respectively, at 100 g/mL fermentation of M.purpureus CWT75. We conclude that the fermentation of M.purpureus CWT75 has a significant anti-invasion and anti-migration activity in hepatoma cells, and exerts anti-oxidative activity in liver cells.

並列關鍵字

Monascus purpureus CWT715 ； anti-cancer ； anti-metastasis ； antioxidants activity

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