Attenuation of LDHA expression in cancer cells leads to redox-dependent alterations in cytoskeletal structure and cell migration
Attenuation of LDHA expression in cancer cells leads to redox-dependent alterations in cytoskeletal structure and cell migration
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DOI：
10.1016/j.canlet.2013.03.034
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发表时间：
2013-09-28
期刊：
影响因子：
9.7
通讯作者：
Cumming, Robert C.
中科院分区：
文献类型：
--
作者：
Arseneault, Robert;Chien, Andrew;Cumming, Robert C.
有氧糖酵解，即使在氧气存在的情况下也优先使用糖酵解来满足细胞代谢需求，是癌症的一个几乎普遍的特征。这种独特的代谢类型被认为可以保护癌细胞免受线粒体中产生的活性氧（ROS）的破坏。通过对癌细胞株MDA-MB-435的研究表明，shRNA介导的乳酸脱氢酶A (LDHA)（有氧糖酵解的关键介质）的敲低导致线粒体ROS产生升高，并伴随细胞增殖和运动性下降。通过氧化还原2D-PAGE和质谱法鉴定了受氧化应激影响的与LDHA敲低相关的氧化还原敏感蛋白。特别是，原肌球蛋白（Tm）亚型Tm4、Tm5NM1和Tm5NM5，这些参与细胞迁移和细胞骨架动力学的蛋白质，在LDHA敲除细胞中表现出二硫键结合和与笔核肌动蛋白聚集体共定位的变化。相反，用巯基抗氧化剂n -乙酰半胱氨酸处理促进Tms向皮质肌动蛋白微丝的重新定位，并部分挽救了与LDHA表达减弱相关的迁移缺陷。这些结果表明，有氧糖酵解和线粒体ROS产生的减少创造了有利于细胞骨架重塑的环境；与癌症相关的高细胞运动性的关键事件。2013爱思唯尔爱尔兰有限公司版权所有。
Aerobic glycolysis, the preferential use of glycolysis even in the presence of oxygen to meet cellular metabolic demands, is a near universal feature of cancer. This unique type of metabolism is thought to protect cancer cells from damaging reactive oxygen species (ROS) produced in the mitochondria. Using the cancer cell line MDA-MB-435 it is shown that shRNA mediated knockdown of lactate dehydrogenase A (LDHA), a key mediator of aerobic glycolysis, results in elevated mitochondrial ROS production and a concomitant decrease in cell proliferation and motility. Redox-sensitive proteins affected by oxidative stress associated with LDHA knockdown were identified by Redox 2D-PAGE and mass spectrometry. In particular, tropomyosin (Tm) isoforms Tm4, Tm5NM1 and Tm5NM5, proteins involved in cell migration and cytoskeletal dynamics, exhibited changes in disulfide bonding and co-localized with pen-nuclear actin aggregates in LDHA knockdown cells. In contrast, treatment with the thiol-based antioxidant N-acetylcysteine promoted the relocalization of Tms to cortical actin microfilaments and partially rescued the migration defects associated with attenuated LDHA expression. These results suggest that aerobic glycolysis and reduced mitochondrial ROS production create an environment conducive to cytoskeletal remodeling; key events linked to the high cell motility associated with cancer. (C) 2013 Elsevier Ireland Ltd. All rights reserved.