EFFECT OF THAI SARAPHI FLOWER EXTRACTS ON WT1 AND BCR/ABL PROTEIN EXPRESSION IN LEUKEMIC CELL LINES.

Rungkarn Sangkaruk, Methee Rungrojsakul, Singkome Tima, Songyot Anuchapreeda
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引用次数: 13

Abstract

Background: Saraphi (Mammea siamensis) is a Thai traditional herb. In this study, the cytotoxic effects of crude ethanolic and fractional extracts including hexane, ethyl acetate, and methanol fractions from M. siamensis flowers were investigated in order to determine their effect on WT1 expression in Molt4 and K562 cells and Bcr/Abl expression in K562 cells.

Materials and methods: The flowers of M. siamensis were extracted using ethanol. The ethanol flower extract was further fractionated with hexane, ethyl acetate, and methanol. Cytotoxic effects were measured by the MTT assay. Bcr/Abl and WT1 protein levels after treatments were determined by Western blotting. The total cell number was determined via the typan blue exclusion method.

Results: The hexane fraction showed the strongest cytotoxic activity on Molt4 and K562 cells, with IC50 values of 2.6 and 77.6 μg/ml, respectively. The hexane extract decreased Bcr/Abl protein expression in K562 cells by 74.6% and WT1 protein expressions in Molt4 and K562 cells by 68.4 and 72.1%, respectively. Total cell numbers were decreased by 66.2 and 48.7% in Molt4 and K562 cells, respectively. Mammea E/BB (main active compound) significantly decreased both Bcr/Abl and WTlprotein expressions by 75 and 49.5%, respectively when compared to vehicle control.

Conclusion: The hexane fraction from M. siamensis flowers inhibited cell proliferation via the suppression of WT1 expression in Molt4 and K562 cells and Bcr/Abl expression in K562 cells. The active compound may be mammea E/BB. Extracts from M. siamensis flowers show promise as naturally occurring anti-cancer drugs.

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泰国沙拉花提取物对白血病细胞wt1和bcr / abl蛋白表达的影响。
背景:萨拉菲(Mammea siamensis)是一种泰国传统草药。本研究研究了暹罗花粗乙醇和部分提取物(包括己烷、乙酸乙酯和甲醇)的细胞毒作用,以确定它们对Molt4和K562细胞中WT1表达和K562细胞中Bcr/Abl表达的影响。材料与方法:采用乙醇提取法提取连翘花。用正己烷、乙酸乙酯和甲醇进一步对乙醇花提取物进行分馏。用MTT法测定细胞毒作用。Western blotting检测处理后Bcr/Abl和WT1蛋白水平。通过typan blue排除法测定细胞总数。结果:正己烷部位对Molt4和K562细胞的细胞毒活性最强,IC50值分别为2.6和77.6 μg/ml。正己烷提取物使K562细胞Bcr/Abl蛋白表达量降低74.6%,使Molt4和K562细胞WT1蛋白表达量分别降低68.4和72.1%。Molt4和K562细胞总细胞数分别减少66.2和48.7%。与对照相比,Mammea E/BB(主要活性化合物)显著降低了Bcr/Abl和WTlprotein的表达量,分别降低了75%和49.5%。结论:暹罗花己烷组分通过抑制Molt4和K562细胞中WT1的表达以及K562细胞中Bcr/Abl的表达来抑制细胞增殖。活性化合物可能是哺乳类E/BB。连翘花提取物有望成为天然的抗癌药物。
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