Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
Department of Materials and Life Sciences, Faculty of Science and Technology, Sophia University, Tokyo 102-8554, Japan.
Molecules. 2020 Mar 12;25(6):1300. doi: 10.3390/molecules25061300.
Kaffir lime () is a plant member of family Rutaceae, and its leaves are commonly used in folk medicine. The present study explores antileukemic effects of the extracts and purified active compounds from the leaves. The antileukemic activity was investigated via inhibition of Wilms' tumor 1 (WT1), which is a protein that involves in leukemic cell proliferation. In addition, the compounds were investigated for their effects on gene expression using real time RT-PCR and Western blotting. Cell cycle arrest and total cell number were investigated using flow cytometry and trypan blue exclusion method, respectively. The results demonstrated that the hexane fractionated extract had the greatest inhibitory effect on gene expression of many leukemic cell lines and significantly decreased WT1 protein levels of K562 cells (representative of the leukemic cells), in a dose- and time-dependent manner. Subfraction No. 9 (F9) after partial purification of hexane fractioned extract showed the highest suppression on WT1 protein and suppressed cell cycle at G2/M. The organic compounds were isolated from F9 and identified as phytol and lupeol. The bioassays confirmed antiproliferative activities of natural products phytol and lupeol. The results demonstrated anticancer activity of the isolated phytol and lupeol to decrease leukemic cell proliferation.
酸橙()是芸香科植物的一员,其叶子常用于民间医学。本研究探讨了该植物叶子的提取物和纯化活性化合物的抗白血病作用。通过抑制 Wilms 瘤 1(WT1)来研究抗白血病活性,WT1 是一种参与白血病细胞增殖的蛋白质。此外,还通过实时 RT-PCR 和 Western blot 法研究了化合物对基因表达的影响。通过流式细胞术和台盼蓝排除法分别研究细胞周期停滞和总细胞数。结果表明,己烷分馏提取物对多种白血病细胞系的基因表达具有最大的抑制作用,并显著降低 K562 细胞(代表白血病细胞)的 WT1 蛋白水平,呈剂量和时间依赖性。己烷分馏提取物部分纯化后的亚馏分 9(F9)对 WT1 蛋白的抑制作用最高,并抑制细胞周期进入 G2/M 期。从 F9 中分离出的有机化合物被鉴定为植醇和羽扇豆醇。生物测定证实了天然产物植醇和羽扇豆醇的增殖抑制活性。结果表明,分离得到的植醇和羽扇豆醇具有抑制白血病细胞增殖的抗癌活性。