十字花科蔬菜衍生物與IL-6對人類胃腺癌細胞程式凋亡的影響

Effects of compunds derived from cruciferous vegetable and IL-6 on apoptosis in gastric cancer cells

黃素華(Su-Hua Huang)

指導教授 ： 陳玉華

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(2003年)

摘要

本研究先觀察胃癌患者術前與術後體內的發炎媒介物質 Interleukin-6 (IL-6)及Nitric oxide (NO)濃度改變的情形，以探討之間的關係。血清分析的結果，胃癌患者術後的第五天IL-6濃度已回復至正常值，雖IgA和IgG尚保持在高濃度的現象(p<0.05)，但仍在正常值的範圍，表示術後合併併發症危險度已獲得降低的可能性。NO的半衰期相當短，於術後第五天則無法觀察到改變的情形。另外，主要以人類胃腺癌細胞 (human gastric adenocarcinoma cell line, AGS)作為體外實驗模式，探討十字花科蔬菜衍生物中的phenylethyl isothiocyanate(PEITC)與indole-3-carbinol(I3C)對於胃腺癌細胞凋亡(apoptosis)的影響。控制組為AGS細胞於0.5% ethanol之培養液中，實驗組分別添加250 μM I3C、10μM PEITC、50 ng/ml IL-6、I3C+IL-6、PEITC+IL-6、I3C+PEITC共七組，結果顯示，雖PEITC有誘導出較多的NO，但不具統計上的意義，I3C和IL-6組則不具有誘導NO的生成。於MTS細胞毒性分析結果顯示，PEITC與I3C可以明顯降低AGS細胞的存活率(p<0.05)。以Flow cytometry分析細胞週期結果顯示，與細胞中添加PEITC(5∼10 μM)24小時後，隨著劑量的增加抑制作用也增強，然不同濃度對於AGS細胞抑制分別有不同的影響，其中以高濃度10 μM的PEITC表現出AGS細胞增加subG0(apoptotic cell)的百分比，其它濃度(5∼7.5 μM)的PEITC則增加G2/M的百分比。250 μM的I3C及添加50 ng/ml的IL-6細胞表現出僅可少許增加細胞中apoptotic cell數目的百分比，且細胞的堆積是在G2/M的百分比。以Annexin Ⅴ-FITC雙染分析，結果顯示，高濃度10 μM的PEITC表現出apoptotic cell (於LR象限)的推積較多的細胞百分比(p<0.05)，結果認為，十字花科蔬菜衍生物中的PEITC與I3C，可以降低AGS細胞的存活率，10 μM 的PEITC可藉由誘導AGS細胞發生apoptosis的現象，進而抑制AGS細胞的生長，而250 μM的I3C對於抑制AGS細胞的生長並非藉由apoptosis的作用機制。

關鍵字

細胞凋亡 ； AGS ； phenylethyl isothiocyanate ； Indole-3-carbinol ； 十字花科蔬菜

並列摘要

As previous study showed that serum interleukin-6(IL-6) and nitric oxide(NO) levels were higher in tumor tissues. This study showed serum concentrations of IL-6 and NO between pre-operation 24 hours and post operation on the day 5th of gastric cancer patients did not significantly differ. Serum IgA and IgG were significantly different(p<0.05). Lots of papers indicated phenylethyl isothiocyanate (PEITC) and indole-3-carbinol (I3C), compounds derivatived from cruciferous vegetable, were phytochemicals which may be responsible for the prevention of many types of cancer. In vitro model, we studied the effects of 10 μM PEITC, 250 μM I3C, 50 ng/ml IL-6 on induction of apoptosis in human gastric adenocarcinoma cells (AGS). Results indicated that NO level in groups of PEITC+LPS and PEITC+IL-6+LPS were higher than other groups. Cell toxicity as measured by MTS assay showed that PEITC and I3C treated groups significantly decreased cell viability. Cell apoptosis as measured by flow cytometry assay with PI analysis showed 10 μM PEITC group increased subG0 (apoptosis) percentage than 5∼7.5 μM PEITC groups. Annexin V-FITC analysis also showed 10mM PEITC group increased apoptosis cell percentage. Therefore, these results suggest that 10 μM PEITC can decrease AGS cell viability throughout apoptosis machanism, but 250 μM I3C did not.

並列關鍵字

apoptosis ； AGS ； phenylethyl isothiocyanate ； Indole-3-carbinol ； cruciferous vegetable derivatives

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