Homoharringtonine targets Smad3 and TGF-β pathway to inhibit the proliferation of acute myeloid leukemia cells.
Homoharringtonine targets Smad3 and TGF-β pathway to inhibit the proliferation of acute myeloid leukemia cells.
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高三尖杉酯碱靶向Smad3和TGF-β通路抑制急性髓系白血病细胞增殖
DOI：
10.18632/oncotarget.16956
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发表时间：
2017-06-20
期刊：
影响因子：
--
通讯作者：
Jin J
中科院分区：
文献类型：
--
作者：
Chen J;Mu Q;Li X;Yin X;Yu M;Jin J;Li C;Zhou Y;Zhou J;Suo S;Lu D;Jin J
高三尖杉酯碱（Homoharringtonine，HHT）在我国长期广泛应用于急性髓细胞白血病（acute myeloid leukemia，AML）的治疗，临床疗效显著，但其作用机制尚不清楚。本研究的目的是通过虚拟筛选的方法筛选HHT可能的作用靶点，并通过细胞实验进行验证。使用包括Autodock、Python和MGL工具的软件，其中HHT为配体，来自PI 3 K-Akt通路、Jak-stat通路、TGF-β通路和NK-κB通路的蛋白质为受体。培养人AML细胞系U937、KG-1、THP-1作为实验细胞系。MTT法检测细胞增殖，流式细胞仪检测HHT作用后细胞凋亡和细胞周期阻滞，Western blotting检测蛋白水平变化，病毒shRNA转染抑制候选蛋白表达，病毒mRNA转染过表达。虚拟筛选结果显示，TGF-β途径中的smad 3可能是HHT结合的候选者。在AML细胞系U937和KG-1中，HHT可诱导Smad 3的Ser 423/425磷酸化，该磷酸化随后可激活TGF-β通路，导致U937细胞周期停滞于G1期，KG-1细胞凋亡，Smad 3的敲低可降低U937细胞对HHT的敏感性，并且Smad 3的过表达可以在两种细胞系中重新建立敏感性。结论：Smad 3可能是HHT的靶蛋白，在HHT的作用机制中起重要作用。
Homoharringtonine (HHT) has long and widely been used in China for the treatment of acute myeloid leukemia (AML), the clinical therapeutic effect is significant but the working mechanism is poorly understood. The purpose of this study is to screen the possible target for HHT with virtual screening and verify the findings by cell experiments. Software including Autodock, Python, and MGL tools were used, with HHT being the ligand and proteins from PI3K-Akt pathway, Jak-stat pathway, TGF-β pathway and NK-κB pathway as the receptors. Human AML cell lines including U937, KG-1, THP-1 were cultured and used as the experiment cell lines. MTT assay was used for proliferation detection, flowcytometry was used to detect apoptosis and cell cycle arrest upon HHT functioning, western blotting was used to detect the protein level changes, viral shRNA transfection was used to suppress the expression level of the target protein candidate, and viral mRNA transfection was used for over-expression. Virtual screening revealed that smad3 from TGF-β pathway might be the candidate for HHT binding. In AML cell line U937 and KG-1, HHT can induce the Ser423/425 phosphorylation of smad3, and this phosphorylation can subsequently activate the TGF-β pathway, causing cell cycle arrest at G1 phase in U937 cells and apoptosis in KG-1 cells, knockdown of smad3 can impair the sensitivity of U937 cell to HHT, and over-expression of smad3 can re-establish the sensitivity in both cell lines. We conclude that smad3 is the probable target protein of HHT and plays an important role in the functioning mechanism of HHT.
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影响因子：
8.8
作者：
通讯作者：
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影响因子：
45.3
作者：
Kantarjian, HM;Talpaz, M;O'Brien, S
通讯作者：
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DOI：
10.1038/nrc3500
发表时间：
2013-05
期刊：
Nature reviews. Cancer
影响因子：
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作者：
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通讯作者：
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DOI：
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发表时间：
1977-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
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作者：
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通讯作者：
VAZQUEZ, D
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DOI：
10.1038/nrm3434
发表时间：
2012-10
期刊：
Nature reviews. Molecular cell biology
影响因子：
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