番杏( Tetragonia tetragonoides (PALL.) KUNTZE)粗抽物誘導人類肝癌細胞株Hep G2細胞週期停滯及細胞凋亡之分子機轉;The Mechanism of crude extract of Tetragonia tetragonoides induced cell cycle arrest and apoptosis in human hepatoblastoma Hep G2 cell lines

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Title:	番杏( Tetragonia tetragonoides (PALL.) KUNTZE)粗抽物誘導人類肝癌細胞株Hep G2細胞週期停滯及細胞凋亡之分子機轉;The Mechanism of crude extract of Tetragonia tetragonoides induced cell cycle arrest and apoptosis in human hepatoblastoma Hep G2 cell lines
Authors:	楊鈞隆;Jiun-Long Yang
Contributors:	中國醫藥大學：中國藥學研究所博士班
Keywords:	番杏;肝癌;細胞凋亡;一氧化氮;西方點墨法;Tetragonia tetragonoides;Hep G2 cell line;Apoptosis;cell cycle;Chinese herbal medicine
Date:	2006-05-29
Issue Date:	2009-08-11 10:25:33 (UTC+8)
Abstract:	番杏（Tetragonia tetragonoides (PALL.) KUNTZE）是一種常用於抗菌、抗腸胃潰瘍的民間中草藥，甚至有時用於治療胃癌、肺癌、肝癌及子宮頸癌，其抗癌的相關機轉並未被充分了解，本研究是探討番杏之50％乙醇粗抽物的抗癌活性機轉。結果發現，番杏之50％酒精萃取物對於人類肝癌細胞株Hep G2導致細胞凋亡，能夠明顯抑制細胞的增生及活動，型態學上觀察這些細胞，可發現凋亡的細胞細胞膜呈現模糊不清，細胞質空泡化，部分細胞甚至變淡死亡，在DNA電泳分析方面可以清楚的見到典型的DNA ladder，細胞內ROS、鈣離子上升，細胞內粒線體膜電位下降。在細胞週期方面，可發現到明顯的停滯在G2/M期，並且能誘導sub-G1 peak的產生，在一氧化氮測定方面，發現能誘導一氧化氮（Nitric Oxide）的產生。以西方點墨法分析蛋白質的表現方面，可以發現凋亡蛋白up-regulation的有PARP、Endo-G、AIF、caspase 12、GADD 153、cytochrome C、Fas、Fas-L、BAX、Bcl-Xs，down-regulation的有NFκB-p65、NFκB-p50、Bcl-2、Bcl-Xl，抑制細胞週期相關蛋白up-regulation的有Chk 1、Wee-1、p21、p27，down-regulation的有CDC 2、CDC25C、cyclin B1，不變的有Chk 2，清除ROS的酵素相關蛋白Mn-SOD為先上升後下降，Cu/Zn-SOD為上升，catalase則是下降、在Nitric oxide相關蛋白iNOS、Nitrotyrosine都有上升，細胞活動轉移相關蛋白MMP-2及MMP-9也有明顯下降，在caspase活性測定也有明顯的活性表現上升現象。其結果可發現番杏的抗癌機轉主要可能是經由ROS的產生、造成細胞的壓力，間接引動caspase活化，造成細胞的傷害。 Herbal medicine has been utilized to treat a variety of diseases, including cancer. Tetragonia tetragonoides (PALL.) KUNTZE (Aizoaceae) is commonly used in Chinese herbal medicine to treat patients with gastrointestinal disorders, especially in stomach and esophageal cancer. The present study, it is to evaluate the effect of the cytotoxic activity on human heptoma Hep G2 cells form extract of T. tetragonoides. Viability assays for extracts of T. tetragonoides showed that it had dose-dependent effect on Hep G2 human cancer cells. The in-vitro cytotoxic effects were investigated by cell cycle analysis and determination of apoptotic DNA fragmentation in Hep G2 cells. The cell cycle analysis showed that extracts of T. tetragonoides induced a significant increase in the number of cells in G2/M phase. The extract of T. tetragonoides -induced chromatin changes and apoptosis of Hep G2 cells also were confirmed by DAPI and PI staining that were measured by fluorescence microscopy and flow cytometry. These results promoted us to further evaluate apoptosis associated proteins and elucidate the possible signal pathway in Hep G2 cells by the extract of T. tetragonoides.
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