Synergistic interactions between HDAC and sirtuin inhibitors in human leukemia cells.
Synergistic interactions between HDAC and sirtuin inhibitors in human leukemia cells.
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DOI：
10.1371/journal.pone.0022739
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Nencioni A
中科院分区：
文献类型：
--
作者：
Cea M;Soncini D;Fruscione F;Raffaghello L;Garuti A;Emionite L;Moran E;Magnone M;Zoppoli G;Reverberi D;Caffa I;Salis A;Cagnetta A;Bergamaschi M;Casciaro S;Pierri I;Damonte G;Ansaldi F;Gobbi M;Pistoia V;Ballestrero A;Patrone F;Bruzzone S;Nencioni A
组蛋白脱乙酰酶(HDAC)活性异常在人类白血病中很常见。然而，尽管经典的、非依赖NAD+的HDAC是一个既定的治疗靶点，但依赖于NAD+的HDAC(Sirtuins)在白血病治疗中的相关性仍不清楚。在这里，我们评估了sirtuin抑制剂和降低NAD+的药物FK866单独以及与传统的HDAC抑制剂联合使用的抗白血病活性。原代白血病细胞、白血病细胞系、健康白细胞和造血祖细胞分别用sirtuin抑制剂(sirtinol，cambinol，EX527)和FK866处理，加或不加HDAC抑制剂丙戊酸、丁酸钠和伏立诺。细胞死亡通过碘化丙啶细胞染色和随后的流式细胞术进行量化。用FITC-Annexin-V/碘化丙啶染色或TMRE染色后进行流式细胞仪分析，并通过检测caspase3/7活性来监测细胞的凋亡诱导。用流式细胞仪和免疫印迹法检测细胞内Bax的表达。用酶循环法测定细胞内NAD+水平。经逆转录病毒转导后，bax基因过表达。BAX和SIRT1被RNA干扰沉默。Sirtuin抑制剂和FK866协同增强白血病细胞中的HDAC抑制剂活性，但在健康白细胞和造血祖细胞中不能。在白血病细胞中，HDAC抑制剂被发现诱导Bax上调，Bax是一种促凋亡的Bcl2家族成员，其易位通常被SIRT1阻止。结果，白血病细胞对sirtuin抑制剂诱导的细胞凋亡变得敏感。综上所述，NAD+非依赖性HDAC和sirtuins在白血病细胞中协同作用以避免细胞凋亡。将sirtuin与HDAC抑制剂联合使用可产生协同抗白血病活性，可用于治疗。
Aberrant histone deacetylase (HDAC) activity is frequent in human leukemias. However, while classical, NAD+-independent HDACs are an established therapeutic target, the relevance of NAD+-dependent HDACs (sirtuins) in leukemia treatment remains unclear. Here, we assessed the antileukemic activity of sirtuin inhibitors and of the NAD+-lowering drug FK866, alone and in combination with traditional HDAC inhibitors. Primary leukemia cells, leukemia cell lines, healthy leukocytes and hematopoietic progenitors were treated with sirtuin inhibitors (sirtinol, cambinol, EX527) and with FK866, with or without addition of the HDAC inhibitors valproic acid, sodium butyrate, and vorinostat. Cell death was quantified by propidium iodide cell staining and subsequent flow-cytometry. Apoptosis induction was monitored by cell staining with FITC-Annexin-V/propidium iodide or with TMRE followed by flow-cytometric analysis, and by measuring caspase3/7 activity. Intracellular Bax was detected by flow-cytometry and western blotting. Cellular NAD+ levels were measured by enzymatic cycling assays. Bax was overexpressed by retroviral transduction. Bax and SIRT1 were silenced by RNA-interference. Sirtuin inhibitors and FK866 synergistically enhanced HDAC inhibitor activity in leukemia cells, but not in healthy leukocytes and hematopoietic progenitors. In leukemia cells, HDAC inhibitors were found to induce upregulation of Bax, a pro-apoptotic Bcl2 family-member whose translocation to mitochondria is normally prevented by SIRT1. As a result, leukemia cells become sensitized to sirtuin inhibitor-induced apoptosis. In conclusion, NAD+-independent HDACs and sirtuins cooperate in leukemia cells to avoid apoptosis. Combining sirtuin with HDAC inhibitors results in synergistic antileukemic activity that could be therapeutically exploited.
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3.4
作者：
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通讯作者：
Hanauske, Axel-Rainer
影响因子：
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Nencioni A
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作者：
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通讯作者：
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