黃豆皂素粗萃取物抑制人類結腸癌 WiDr 細胞生長機制之探討

Growth inhibitory mechanisms of extracted crude soybean saponins on human colon cancer WiDr cells

蔡政諭(Cheng-Yuh Tsai)

指導教授 ： 林士祥

臺北醫學大學/公共衛生暨營養學院/保健營養學研究所/碩士(2005年)

摘要

癌症連續二十年為國人十大死亡原因第一位，其中結直腸癌為成人癌症死因排名第三位，在西方國家中罹患結腸癌的機率高於東亞國家，原因可能為東亞國家較西方國家攝取較多的黃豆及豆製品。而黃豆中皂素有抑制結腸癌的效果，因此本實驗將探討其和結腸癌之關係。實驗設計及方法：選用結腸癌細胞 WiDr 細胞株加入不同濃度萃取的黃豆皂素分別為 150 ppm、300 ppm、600 ppm、1200 ppm 觀察其對細胞的生長、alkaline phosphatase (AP)、protein kinase C (PKC) 的活性 及P53、c-Fos、c-Jun 蛋白質表現的影響，並利用掃描式電子顯微鏡 (SEM) 及穿透式電子顯微鏡 (TEM) 觀察其對細胞形態的影響。結果發現 300 ppm、600 ppm、1200 ppm 濃度的黃豆皂素粗萃取物可抑制 WiDr 細胞的生長，改變細胞形態，使細胞質產生空泡，並且抑制由 12-O-tetradecanol phorbol 13-acetate 誘導之 PKC 活性。600 ppm與 1200 ppm 濃度的黃豆皂素粗萃取物可增加 AP 的活性。因此推測黃豆皂素粗萃取物可以促使 Widr 細胞分化，抑制細胞增生，並誘導 type II autophagic death 而抑制細胞的生長。

關鍵字

黃豆皂素 ； alkaline phosphatase ； protein kinase C ； P53 ； c-Jun ； c-Fos

並列摘要

Among the cancers that cause most deaths in adults, colon cancer is ranked the third. Because people in the Asian countries intake more soybeans and soy-based products than those in the Western countries, the incidence of colon cancer is lower. The objective of this study was to investigate the effect of extracted crude soybean saponins on human colon cancer cells. In this study, the culture of WiDr (human colon cancer cells) was treated with 150 ppm, 300 ppm, 600 ppm, 1200 ppm of saponins to determine the effect on cell growth, morphology, alkaline phosphatase (AP) activity, and protein kinase C (PKC) activity, and P53, c-Fos and c-Jun expression of colon cells. Results indicated that soybean saponins decreased cell growth in a dose-dependent manner, and pre-treatment of cells with saponins significantly suppressed the 12-O-tetradecanol phorbol 13-acetate-stimulated PKC activity.Treating Cells with 600 and 1200 ppm of saponins significantly increased AP activity. Cells treated with saponins developed cytoplasmic vesicles and wrinkled plasma membrane. However, the effects of saponins on P53, c-Fos and c-Jun expression were not significant. In conclusion, soybean saponins interacted with cell membranes, suppressed PKC activation and induced diffrtrntiation, and induce type II autophagic death, which possibly mediate the growth inhibition of tumor cells.

並列關鍵字

soybean saponins ； alkaline phosphatase ； protein kinase C ； P53 ； c-Jun ； c-Fos

參考文獻

Akao T, Kida H, Kanaoka M, Hattori M and Kobashi K (1998) Intestinal bacterial hydrolysis is required for the appearance of compound K in rat plasma after oral administration of ginsenosides Rb1 from Panax ginseng. J. Pharm. Pharmacol. 50:1155-1160.

Angel P and Karin M. (1991) The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta. 1072(2-3):129-157.

Bae EA, Han MJ, Choo MK, Park SY and Kim DH (2002) Metabolism of 20(S)- and 20(R)-ginsenoside Rg3 by human Intestinal bacteria and its relation to in vitro biological activities. Biol. Pharm. Bull. 25:58-63.

Baehrecke EH. (2002) How death shapes life during development. Nat Rev Mol Cell Biol. 3(10):779-787.

Basson MD, Hong F and Emenaker NJ. (1995) Specific modulation of intestinal epithelial brush border enzyme expression by a phorbol ester. J Surg Res. 59(1):121-126.