Department of Hematology, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, China.
Department of General Practice, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, China.
Life Sci. 2019 Jun 15;227:153-165. doi: 10.1016/j.lfs.2019.04.034. Epub 2019 Apr 17.
Alantolactone (ALT) is active component of natural product Inula helenium with a lot of pharmacological effects, including anti-tumor effect. The present work aimed to explore the antitumor effect of ALT in B cell acute lymphoblastic leukemia (B-ALL).
B-ALL cells were treated with various concentrations of ALT, and then trypan blue assay, Annexin V/PI staining assay, PI staining assay, western blot analysis were employed to measure the effect of ALT on viability, apoptosis and cell cycle in B-ALL cells. In addition, a synthetic bioinformatics method was used to predict the underlying mechanism of antitumor effect of ALT. Then Reactive Oxygen Species (ROS) probe Dihydroethidium (DHE) and 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) were used to detect accumulation of cellular ROS. Meanwhile, DNA damage was identified by 8-oxoG, p-ATM1987, γ-H2AX and comet assay. In addition, activity of glutathione reductase (GR), thioredoxin reductase (TrxR) and catalase were measured and overexpressed in SEM and RS4;11 cells to study the inhibition on these enzymes. Finally, B-ALL NOD-SCID mouse model was used to test its performance in vivo.
ALT showed good antitumor effect in B-ALL in vivo and in vitro through inducing ROS overload, which led to DNA damage. In addition, we found ROS overload caused by ALT was due to its direct inhibition on reductase.
We found that ALT, a natural product, showing a promising tactic in the therapy of B-ALL by targeting ROS pathway.
土木香内酯（ALT）是旋覆花属天然产物的活性成分，具有多种药理作用，包括抗肿瘤作用。本研究旨在探讨 ALT 对 B 细胞急性淋巴细胞白血病（B-ALL）的抗肿瘤作用。
用不同浓度的 ALT 处理 B-ALL 细胞，然后采用台盼蓝染色法、Annexin V/PI 染色法、PI 染色法、Western blot 分析检测 ALT 对 B-ALL 细胞活力、凋亡和细胞周期的影响。此外，采用合成生物信息学方法预测 ALT 抗肿瘤作用的潜在机制。然后用活性氧（ROS）探针二氢乙啶（DHE）和 2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）检测细胞内 ROS 的积累。同时，通过 8-氧鸟嘌呤（8-oxoG）、p-ATM1987、γ-H2AX 和彗星试验鉴定 DNA 损伤。此外，还测量并过表达 SEM 和 RS4;11 细胞中的谷胱甘肽还原酶（GR）、硫氧还蛋白还原酶（TrxR）和过氧化氢酶的活性，以研究对这些酶的抑制作用。最后，在 B-ALL NOD-SCID 小鼠模型中测试其体内性能。
ALT 通过诱导 ROS 过载导致 DNA 损伤，在体内和体外均显示出良好的抗肿瘤作用。此外，我们发现 ALT 引起的 ROS 过载是由于其对还原酶的直接抑制。
我们发现，天然产物 ALT 通过靶向 ROS 通路，为 B-ALL 的治疗提供了一种有前途的策略。
