使用洛神葉萃取物在人類黑色素瘤及前列腺癌細胞的抗癌效果

Anti-Cancer Effect of Hibiscus sabdariffa Leaf Extract (HLP and HLE) on Human Melanoma and Human Prostate Cancer Cells

邱軍棠(Chun-Tang Chiu)

指導教授 ： 周芬碧 ； 陳璟賢

中山醫學大學/醫學院/生化暨生物科技研究所/博士(2015年)

https://doi.org/10.6834/CSMU.2015.00017

摘要

近年來已有許多研究指出，洛神葉具有降血脂、降血糖、抗氧化與促進腫瘤細胞凋亡的作用。然而，目前對於取自洛神葉的萃取物質，包含洛神葉多酚萃取物(Hibiscus sabdariffa leaf polyphenolic extract ; HLP)與洛神葉萃取物(Hibiscus sabdariffa leaf extract; HLE)等的詳細抗癌分子機制，卻所知有限。因此，本研究主題欲探討Hibiscus sabdariffa leaf polyphenolic extract(HLP)與Hibiscus sabdariffa leaf extract(HLE)抗癌能力的研究。首先，從分析發現，Hibiscus sabdariffa leaf polyphenolic extract(HLP)含有表兒茶素沒食子酸酯(epicatechin gallate ; ECG)與其他多酚物質，再經由細胞凋亡分析(DAPI染色分析、細胞週期分析)與細胞自噬作用分析(AVO染色分析)發現，HLP具有促進人類黑色素瘤A375細胞的細胞凋亡與細胞自噬作用；細胞凋亡方面發現HLP可活化caspases、調控Bcl-2家族蛋白、活化Fas/FasL路徑來促使A375細胞凋亡；細胞自噬方面發現HLP可活化ATG5、Beclin1和LC3-II來促使A375細胞自噬作用。其次，分析Hibiscus sabdariffa leaf extract(HLE)抑制人類前列腺癌LNCaP細胞侵襲能力，經由傷口癒合試驗與細胞侵襲試驗發現，HLE於無毒劑量下具有劑量依賴性抑制LNCaP細胞侵襲的能力，而此分子機制是經由抑制Akt/NF-kB/MMP-9訊息傳遞路徑；最後再由活體動物試驗證實HLE能抑制人類前列腺癌LNCaP細胞生長與抑制Akt/NF-kB/MMP-9訊息傳遞路徑。本研究證實HLP與HLE確實具有抗癌功效，未來可發展為抗癌藥劑。

關鍵字

洛神葉 ； 多酚 ； 表兒茶素沒食子酸酯 ； 細胞凋亡 ； 細胞自噬 ； 轉移 ； Akt/NF-kB/MMP-9訊息路徑

並列摘要

Hibiscus sabdariffa leaf has been previously shown to possess hypoglycemic, hypolipidemic, and antioxidant effects, and induce tumor cell apoptosis. However, the molecular mechanisms involved in the anticancer activity of Hibiscus sabdariffa leaf polyphenolic extract (HLP) and H. sabdariffa leaf extract (HLE) are poorly understood. The object of the study was to examine the anti-cancer potential of HLP and HLE. First, HLP was exhibited to be rich in epicatechin gallate (ECG) and other polyphenols. Apoptotic and autophagic activities of HLP and ECG were further evaluated by DAPI stain, cell cycle analysis, and acidic vascular organelle (AVO) stain. Our results revealed that both of HLP and ECG induced the caspases cleavages, Bcl-2 family proteins regulation, and Fas/FasL activation in human melanoma A375 cells. In addition, HLP could increase the expressions of autophagy-related proteins autophagy-related gene 5 (ATG5), Beclin1 and light chain 3-II (LC3-II), and induce autophagic cell death in A375 cells. Second, we want to examine the anti-invasive potential of HLE. The results of wound-healing assay and in vitro transwell assay revealed that HLE dose-dependently inhibited the migration and invasion of human prostate cancer LNCaP cells under non-cytotoxic concentrations. Molecular data showed all these influences of HLE might be mediated via inhibition of protein kinase B (PKB, also known as Akt)/NF-kB/MMP-9 cascade pathway. Finally, the inhibitory effect of HLE was proven by its inhibition on the growth of LNCaP cells and the expressions of metastasis-related molecular proteins in vivo. These data indicated that the anti-cancer effect of HLP and HLE. HLP and HLE potentially could be developed as an anti-cancer agent.

並列關鍵字

Hibiscus sabdariffa leaf ； Polyphenol ； Epicatechin gallate ； Apoptosis ； Autophagy ； Metastasis ； Akt/NF-kB/MMP-9 cascade pathway

參考文獻

22.Zeestraten EC, Benard A, Reimers MS, Schouten PC, Liefers GJ, van de Velde CJ, Kuppen PJ: The Prognostic Value of the Apoptosis Pathway in Colorectal Cancer: A Review of the Literature on Biomarkers Identified by Immunohistochemistry. Biomark Cancer. 2013; 5: 13-29.

1.Lin HH, Chen JH, Wang CJ. Chemopreventive properties and molecular mechanisms of the bioactive compounds in Hibiscus sabdariffa Linne. Curr Med Chem. 2011; 18: 1245-1254.

3.Lee SH, Jaganath IB, Wang SM, Sekaran SD. Antimetastatic effects of Phyllanthus on human lung (A549) and breast (MCF-7) cancer cell lines. PLoS One. 2011; 6: 6.

4.Lin HH, Chan KC, Sheu JY, Hsuan SW, Wang CJ, Chen JH. Hibiscus sabdariffa leaf induces apoptosis of human prostate cancer cells in vitro and in vivo. Food Chem. 2012; 132: 880-891.

5.Chen JH, Wang CJ, Wang CP, Sheu JY, Lin CL, Lin HH: Hibiscus sabdariffa leaf polyphenolic extract inhibits LDL oxidation and foam cell formation involving up-regulation of LXRα/ABCA1 pathway. Food Chem. 2013; 141(1): 397-406.