- 學位論文
沒食子酸月桂酯對全反式維甲酸誘發人類急性骨髓癌細胞分化及死亡影響
The effect of lauryl gallate on all trans retinoic acid-induced cell differentiation and death in human acute myelogenous leukemia (AML) cells
摘要
目前治療急性骨髓性白血的藥物，主要的作用是促進癌細胞凋亡，及誘導癌細胞分化。儘管對於急性前骨髓性白血病，分化治療有很好的反應，但分化治療反應所引起的副作用，及其它型態的急性前骨髓性白血病，仍無有效的治療方法。因此尋求有效、副作用低的療法是刻不容緩的課題。沒食子酸月桂酯 (lauryl gallate) 是法定的食品添加物，作為抗氧化劑使用，具有抗氧化、抗發炎，及抗腫瘤等生物活性。本研究之目的在探討沒食子酸月桂酯對全反式維他命甲酸 (all-trans retinoi c acid，ATRA)，誘發人類急性骨髓性白血病細胞 HL-60 和 KG-1 之分化及凋亡作用之影響。研究結果顯示，以1 μΜ 反式維他命甲酸合併3 μΜ 沒食子酸月桂酯處理 HL-60和 KG-1細胞，從流式細胞儀分析，明顯看見增加細胞表面分化標記分子 CD14 及 CD11b的表現。 Wright-Giema 染色也顯示了細胞分化的現象，且隨合併劑量增加，具有加乘效果。實驗中我們亦發現處理反式維他命甲酸合併沒食子酸月桂酯，隨著藥物增加，凋亡細胞也增加。進一步利用西方墨點法，探討誘導細胞凋亡之分子機制，發現合併藥物處理會促使，內在途經抗凋亡分子 Bcl-2 及 Mcl-1 表現量減少；另一方面促進凋亡分子 Bak、Bax 及 PUMA的表現增加；外在途經則透過活化TRAIL之DR4、DR5、Fas及Fas-ligand，最後活化caspase-2、-3、-6、-8及-9，誘導細胞凋亡。由本研究之結果得知，全反式維他命甲酸合併沒食子酸月桂酯可誘導人類血癌細胞HL-60和KG-1，往單核球 (Neutrophil) 和顆粒球 (Monocytes) 系列分化，調控 CD14 和 CD11b 分子的表現。此外，全反式維他命甲酸合併沒食子酸月桂酯也可透過調控 Bcl-2家族分子之表現，經由粒腺體活化的途徑，誘導細胞凋亡。因此，推論沒食子酸月桂酯 (LG)，極具潛力開發為與全反式維他命甲酸 (ATRA)合併使用於促進血癌分化及凋亡治療藥物。
並列摘要
Currently, the most common strategy to treatment of acute myeloid leukemia is to induce apoptosis and differentiation of this tumor cells. The addition of differentiation induction therapy with all trans-retinoic acid (ATRA) is now regarded as mandatory in the treatment of acute promyelocytic leukemia (APL) with a good complete remission rate, but some severe side effects mediated by ATRA are challenge to be overcome. Therefore, seek effective drug, and lower side effects of therapy are essential issues for AML treatment. Lauryl ester of gallic acid (lauryl gallate) is commonly used as an antioxidant of food additives. lauryl gallate has anti-inflammatory and anti-cancer effects. This study aimed to explore the effect of lauryl gallate on the reactivity of ATRA in both APL and non-APL AML cells in vitro using human AML cell lines, HL-60 and KG-1. Results showed that 1μM ATRA combined with 3μM lauryl gallate significantly induced differentiation of HL60 and KG-1 cells comfirmed by morphological changes and increase of CD14 and CD11b monocytic surface marker expression. Our data also exhibited that ATRA plus lauryl gallate treatment induced apoptosis in a dose- and time-dependent manner. Data from Western blot analysis indicated that combined treatment with ATRA and lauryl gallate decreased the expression levels of anti-apoptotic molecules Bcl-2 and Mcl-1, and increased the level of proapoptotic molecules PUMA, Bak and Bax. At the meanwhile, caspase -2,-3,-6,-8 and -9 were activated by combined treatment. Over all, our observations showed that ATRA combined with lauryl gallate could induce human AML HL-60 and KG-1 cell toward both neutrophil and monocyte differentiation. In addition, ATRA plus lauryl gallate treatment also triggered AML cell apoptosis through Bcl-2 family molecule-dependent mitochondrion pathway. Based on these findings, we demonstrate that ATRA plus lauryl gallate will have the development potential of anti-cancer combined treatments. These results suggest that lauryl gallate might have the potential to combine used with ATRA as a therapeutic regimen in AML patients.
參考文獻
延伸閱讀
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