十字花科蔬菜衍生物與

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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

中文摘要

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 μ M 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 μ

M 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。

英文摘要

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

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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.

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