Targeting colorectal cancer cells by a novel sphingosine kinase 1 inhibitor PF-543
Targeting colorectal cancer cells by a novel sphingosine kinase 1 inhibitor PF-543
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DOI：
10.1016/j.bbrc.2016.01.053
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发表时间：
2016-02-12
影响因子：
3.1
通讯作者：
Fang, Zheng-yu
中科院分区：
文献类型：
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作者：
Ju, TongFa;Gao, DaQuan;Fang, Zheng-yu
在这项研究中，我们发现一种新的鞘氨醇激酶1(SphK1)抑制剂PF-543对一组已建立的(HCT-116、HT-29和DLD-1)和原代人类结直肠癌(CRC)细胞具有强大的抗增殖和细胞毒作用。其敏感性与结直肠癌细胞中SphK1的表达水平呈负相关。令人惊讶的是，PF-543主要诱导结直肠癌细胞程序性坏死，而不是凋亡。通过乳酸脱氢酶(LDH)释放、线粒体膜电位(MMP)崩溃和线粒体P53-亲环素-D(Cyp-D)络合检测大肠癌细胞的坏死性死亡。相应地，坏死抑制物Necrostatin-1可显著减弱PF-543对结直肠癌细胞的杀伤作用。同时，Cyp-D抑制剂(sanglife hrin A和环孢素A)或shRNA介导的Cyp-D基因敲除显著减轻了PF-543诱导的CRC细胞的坏死性死亡。反之，野生型Cyp-D在HCT-116细胞中的过表达显著增加了对PF-543‘S的敏感性。在体内，静脉注射PF-543可显著抑制严重联合免疫缺陷(SCID)小鼠的HCT-116移植瘤生长，显著提高小鼠的存活率。当PF-543与Cyp-D抑制剂环孢素A联合使用时，其体内活性大大减弱。我们的结果表明，PF-543在体外和体内都具有很强的抗结直肠癌活性。线粒体程序性坏死途径可能是PF-543‘S作用于大肠癌细胞的关键机制。(C)2016 Elsevier Inc.保留所有权利。
In this study, we showed that PF-543, a novel sphingosine kinase 1 (SphK1) inhibitor, exerted potent anti-proliferative and cytotoxic effects against a panel of established (HCT-116, HT-29 and DLD-1) and primary human colorectal cancer (CRC) cells. Its sensitivity was negatively associated with SphK1 expression level in the CRC cells. Surprisingly, PF-543 mainly induced programmed necrosis, but not apoptosis, in the CRC cells. CRC cell necrotic death was detected by lactate dehydrogenase (LDH) release, mitochondrial membrane potential (MMP) collapse and mitochondrial P53-cyclophilin-D (Cyp-D) complexation. Correspondingly, the necrosis inhibitor necrostatin-1 largely attenuated PF-543-induced cytotoxicity against CRC cells. Meanwhile, the Cyp-D inhibitors (sanglifehrin A and cyclosporin A), or shRNA-mediated knockdown of Cyp-D, remarkably alleviated PF-543-induced CRC cell necrotic death. Reversely, over-expression of wild-type Cyp-D in HCT-116 cells significantly increased PF-543's sensitivity. In vivo, PF-543 intravenous injection significantly suppressed HCT-116 xenograft growth in severe combined immunodeficient (SCID) mice, whiling remarkably improving the mice survival. The in vivo activity by PF-543 was largely attenuated when combined with the Cyp-D inhibitor cyclosporin A. Collectively, our results demonstrate that PF-543 exerts potent anti-CRC activity in vitro and in vivo. Mitochondrial programmed necrosis pathway is likely the key mechanism responsible for PF-543's actions in CRC cells. (C) 2016 Elsevier Inc. All rights reserved.