薏苡籽實乙醇萃取物及其次區分層對人類癌細胞生長之影響

Effect of the fractions and sub-fractions of adlay seed ethanolic extracts on the growth of human cancer cells

林靜慧(Jing-Hui Lin)

指導教授 ： 江文章

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2007年)

https://doi.org/10.6342/NTU.2007.02739

摘要

近年來研究指出薏苡籽實萃取物具有抗腫瘤的生理活性。本研究探討薏苡籽實不同部位乙醇萃取物之區分層和次區分層對於人類肝癌 (Hep 3B和Hep G2)、乳癌 (MCF7)、胃癌 (AGS)和子宮頸癌 (HeLa)細胞株生長的影響及可能的作用機制。薏苡籽實不同部位之乙醇萃取物包含薏苡殼 (AHE)、薏苡種皮 (ATE)、薏苡麩皮 (ABE)及精白薏仁 (PAE)。並依極性不同，可將薏苡籽實不同部位之乙醇萃取物 (AHE、ATE、ABE和PAE)區分成正己烷層 (He)、乙酸乙酯層 (Ea)、正丁醇 (Bu)和水層 (Wa)。由MTT分析法結果顯示，AHE-Ea和ATE-Ea對於所有癌細胞生長具有顯著的抑制能力且具有濃度效應，因此更進一步經由管柱層析區分出次區分層。結果顯示AHE-Ea-K、AHE-Ea-L、ATE-Ea-E和ATE-Ea-F次區分層經72小時處理後可抑制婦癌細胞株 (MCF7和HeLa)之細胞存活率，IC50值介於13.7-58.2 μg/mL。由細胞週期分析可知AHE-Ea-K、AHE-Ea-L、ATE-Ea-E和ATE-Ea-F會誘導MCF7之細胞週期停滯於G0/G1期。此外，經AHE-Ea-K和AHE-Ea-L處理後HeLa之細胞週期會停滯於S期，而ATE-Ea-E和ATE-Ea-F則是誘導HeLa之細胞週期停滯於G0/G1期。根據流式細胞儀的檢測結果指出AHE-Ea-K、AHE-Ea-L、ATE-Ea-E和ATE-Ea-F會經由活化caspase-3誘導MCF7和HeLa產生細胞凋亡。綜合以上結果，AHE-Ea-K、AHE-Ea-L、ATE-Ea-E和ATE-Ea-F可經誘導MCF7和HeLa細胞凋亡以抑制其生長。

關鍵字

薏苡籽實 ； 人類乳癌細胞MCF7 ； 人類子宮頸癌細胞HeLa ； 半胱胺酸蛋白酶-3 ； 細胞凋亡 ； MTT分析法 ； 細胞週期

並列摘要

A number of recent studies have shown the physiological activity in anti-tumor effects of adlay extracts. The study was to investigate the effect of fractions and sub-fractions of adlay seed ethanolic extract on the human liver (Hep 3B and Hep G2), breast (MCF7), stomach (AGS) and cervical (HeLa) cancer cell lines and their possible mechanisms. The ethanolic extracts from different parts of adlay seed were named as AHE (hull), ATE (testa), ABE (bran) and PAE (polished adlay). Based on the degree of polarity, these ethanolic extracts (AHE, ATE, ABE and PAE) were separated to hexane layer (He), ethyl acetate layer (Ea), 1-butanol layer (Bu) and water layer (Wa). The results of 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl- tetrazolium bromide (MTT) assay showed that AHE-Ea and ATE-Ea had the significant capacity against growth of all cancer cells and in the dose-dependent manner, so further partitioned into their sub-fractions via column chromatography. The results showed that AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F sub-fractions inhibited the cell viability of gynecological cancer cell lines (MCF7 and HeLa) for 72 hr, and their IC50 values were 13.7-58.2 μg/mL. The results of cell cycle analysis showed that AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F induced MCF7 cell cycle arrest in G0/G1 phase. Furthermore, cell cycle of HeLa arrest in S phase by AHE-Ea-K and AHE-Ea-L, but ATE-Ea-E and ATE-Ea-F induced its cell cycle arrest in G0/G1 phase. According to the results of flow cytometric analysis, it indicated that AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F induced MCF7 and HeLa apoptosis through induction of caspase-3 activity. In conclusion, AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F could be through inducing apoptosis of MCF7 and HeLa cell lines to inhibit their viability.

並列關鍵字

adlay seed ； human breast cancer cell line MCF7 ； human cervical cancer cell line HeLa ； caspase-3 ； apoptosis ； MTT assay ； cell cycle

參考文獻

江文章，徐明麗，蘇瑞斌，龐飛。2000。薏仁加工食品輔助抑制腫瘤功效之評估。醫護科技學刊2:113-122。

余嚴尊。2004。糙薏仁的活性區分物對倉鼠脂質代謝之研究。台灣大學食品科技研究所。博士論文。台北。

施純光。2003。薏仁對大腸癌形成之影響。台灣大學食品科技研究所。博士論文。台北。

夏詩閔。2005。薏苡麩皮及薏苡殼甲醇萃取物對大鼠卵巢顆粒細胞分泌孕酮之效應。台灣大學食品科技研究所。博士論文。台北。

曾怡雯。2003。薏苡籽實萃取物對大鼠萊氏細胞分泌睪固酮的效應。台灣大學食品科技研究所。碩士論文。台北。