Withanolide Metabolites Inhibit PI3K/AKT and MAPK Pro-Survival Pathways and Induce Apoptosis in Acute Myeloid Leukemia Cells.
Withanolide Metabolites Inhibit PI3K/AKT and MAPK Pro-Survival Pathways and Induce Apoptosis in Acute Myeloid Leukemia Cells.
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DOI：
10.3390/biomedicines8090333
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发表时间：
2020-09-06
期刊：
影响因子：
4.7
通讯作者：
Cutillas PR
中科院分区：
文献类型：
--
作者：
Akhtar N;Baig MW;Haq IU;Rajeeve V;Cutillas PR
急性髓性白血病（AML）是一种侵袭性疾病，尽管取得了进展，但其治疗仍然具有挑战性。因此，寻找治疗这种疾病的新药仍然很重要。Withanolides是茄科植物中发现的一组甾体内酯，由于其在不同环境中的抗癌活性而引起人们的潜在兴趣。在这项研究中，我们研究了茄科衍生的withanolides在AML细胞模型中的抗增殖作用和作用模式;这些代谢产物包括分别从洋金花和Withania coagluanse中分离的withametelin（WTH）和Coagulansin A（CoA）。两种睡茄内酯均抑制AML细胞的增殖并诱导细胞死亡，在所测试的AML模型中，WTH比CoA更有效。采用定量无标记蛋白质组学和磷酸化蛋白质组学来确定所研究的睡茄内酯的作用机制。我们在用WTH、CoA或载体对照处理的细胞中鉴定并定量了5269种蛋白质和17，482个磷酸化位点。Withanolides调节参与调节关键细胞过程的蛋白质的表达，包括细胞周期、代谢、信号传导、蛋白质降解和基因表达。针对激酶底物、激酶-激酶关系和经典途径的磷酸化蛋白质组学数据的富集分析表明，睡茄内酯降低了激酶如磷酸肌醇3-激酶（PI 3 K）、蛋白激酶B（PKB;也称为RAC-α丝氨酸/苏氨酸蛋白激酶或AKT），雷帕霉素的哺乳动物靶蛋白（mTOR），细胞外信号调节蛋白激酶1和2（ERK 1/2）和丝氨酸/苏氨酸蛋白激酶A-Raf（ARAF），同时增加DNA修复激酶的活化。这些结果表明，withanadine代谢物在调节致癌促存活和促凋亡信号通路中具有多效性作用，所述促存活和促凋亡信号通路调节凋亡的诱导。通过免疫印迹法证实，作为处理的结果，显示裂解的PARP和半胱天冬酶3、8和9的表达增加。总体而言，我们的研究结果表明，WTH和CoA对AML具有治疗潜力，WTH表现出更有效的作用，应进一步探索。
Acute myeloid leukemia (AML) is an aggressive disease and, despite advances, its treatment remains challenging. Therefore, it remains important to identify new agents for the management of this disease. Withanolides, a group of steroidal lactones found in Solanaceae plants are of potential interest due to their reported anticancer activities in different settings. In this study we investigated the anti-proliferative effects and mode of action of Solanaceae-derived withanolides in AML cell models; these metabolites include withametelin (WTH) and Coagulansin A (CoA) isolated from Datura innoxia and Withania coagluanse, respectively. Both withanolides inhibited the proliferation of AML cells and induced cell death, with WTH being more potent than CoA in the AML models tested. Quantitative label-free proteomics and phosphoproteomics were employed to define the mechanism of action of the studied withanolides. We identified and quantified 5269 proteins and 17,482 phosphosites in cells treated with WTH, CoA or vehicle control. Withanolides modulated the expression of proteins involved in regulating key cellular processes including cell cycle, metabolism, signaling, protein degradation and gene expression. Enrichment analysis of the phosphoproteomics data against kinase substrates, kinase-kinase relationships and canonical pathways showed that the withanolides decreased the activity of kinases such as phosphoinositide 3-kinase (PI3K), protein kinase B (PKB; also known as RAC-alpha serine/threonine-protein kinase or AKT), mammalian target of rapamycin (mTOR), extracellular signal-regulated protein kinase 1 and 2 (ERK1/2) and the serine/threonine-protein kinase A-Raf (ARAF), while increasing the activation of DNA repair kinases. These results indicate that withanolide metabolites have pleiotropic effects in the modulation of oncogenic pro-survival and pro-apoptotic signaling pathways that regulate the induction of apoptosis. Withanolide mediated apoptosis was confirmed by immunoblotting showing increased expression of cleaved PARP and Caspases 3, 8 and 9 as a result of treatment. Overall, our results suggest that WTH and CoA have therapeutic potential against AML with WTH exhibiting more potent effects and should be explored further.
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影响因子：
37.3
作者：
Mondal S;Mandal C;Sangwan R;Chandra S;Mandal C
通讯作者：
Mandal C
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12.8
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Derolf ÅR
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3.5
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8
作者：
Billottet, C.;Grandage, V. L.;Khwaja, A.
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Khwaja, A.
影响因子：
4.3
作者：
Kong, Ying;Nan, Kejun;Yin, Yuxin
通讯作者：
Yin, Yuxin