巴西洋菇(Agaricus blazei)發酵產物乙醇萃取物 抑制人類肝癌細胞株生長與侵犯之研究

Abstract

行政院衛生署公佈94年國人十大死亡原因惡性腫瘤仍然位居第一名，而肝臟疾病在台灣的發生率非常高，例如慢性肝炎、肝硬化與肝癌，因此肝癌往往是主要的死亡原因之一，肝癌在男性惡性腫瘤死亡原因為第一名，在女性則為第二名，如何治療肝癌的發生是本研究之目標。 本研究結果顯示以0-200 μg/mL的巴西洋菇發酵產物乙醇萃取物(ethanolic extract of fermentation product of Agricus blazei, AB-pE)處理人類肝癌細胞株HA22T/VGH及SK-Hep-1 48小時後，細胞生存力會隨著濃度增加而減少，IC50分別為28.7及26.8 μg/mL，再藉由流式細胞儀來分析細胞週期，結果顯示在處理100及200 μg/mL AB-pE會誘導細胞走向細胞凋亡。DNA 片段化（DNA ladder）的結果顯示出處理AB-pE 100及200 μg/mL於HA22T/VGH細胞，有DNA 片段化的表現。進一步利用螢光及流式細胞儀偵測細胞caspase 3的表現，結果顯示出AB-pE可以活化caspase 3而進行細胞凋亡。利用Matrigel invasion assay來檢測AB-pE對於人類肝癌細胞株HA22T/VGH及SK-Hep-1侵犯力的影響，初步結果顯示在AB-pE處理濃度為50 μg/ml且對細胞毒性不大具有抑制細胞侵犯的作用。綜合上述，巴西洋菇乙醇萃取物具有抑制肝癌細胞的生長及侵犯的能力，可作為開發保健食品之食材。

Department of Health, Executive Yuan, R.O.C, reported that leading causes of death malignant neoplasma still in first place. The incidence of liver disease is very high in Taiwan, for example: chronic hepatitis, cirrhosis and hepatocellular carcinoma. Hepatocellular carcinoma is the most cause of death in Taiwan. Hepatocellular carcinoma is at first place in male and at second place in female in Taiwan. Therefore, how to prevent and heal Hepatocellular carcinoma is very importment. In this study, the results show that when treated 0-200 μg/mL of the ethanolic extract of fermentation product of Agricus blazei (AB-pE) after 48 hr on human hepatocellular carcinoma HA22T/VGH and SK-Hep-1 cells, the cell viability is decrease while concentration increasing. The IC50 of HA22T/VGH and SK-Hep-1 are 28.7 μg/mL and 26.8 μg/mL. In DNA fragmentation experiment, when treated 100, 200 μg/mL on HA22T/VGH cells, the DNA will cleave by nuclease and form DNA ladder. Using flow cytometry to detect the cell cycle, the results show that when treated 100 and 200 μg/ml of AB-pE after 48 hr would lead cell toward apoptosis. And then using fluorescence and flow cytometry to detect the activity of caspase 3, the results show that the AB-pE can activate caspase 3 and cause apoptosis. Using Matrigel invasion assay to detect the invasion ability of human hepatocellular carcinoma HA22T/VGH and SK-Hep-1, when treated 50 μg/ml AB-pE, the hepatoma’s invasion activity has decrease. In conclusion, the results show that the ethanolic extract of fermentation product of Agricus blazei (AB-pE) could inhibit the cell proliferation and the invasion activity of human hepatocellular carcinoma HA22T/VGH and SK-Hep-1.