The oxidoreductase CLIC4 is required to maintain mitochondrial function and resistance to exogenous oxidants in breast cancer cells.
The oxidoreductase CLIC4 is required to maintain mitochondrial function and resistance to exogenous oxidants in breast cancer cells.
复制标题
DOI：
10.1016/j.jbc.2022.102275
复制
发表时间：
2022-09
影响因子：
4.8
通讯作者：
Yuspa, Stuart H.
中科院分区：
文献类型：
--
作者：
Al Khamici, Heba;Sanchez, Vanesa C.;Yan, Hualong;Cataisson, Christophe;Michalowski, Aleksandra M.;Yang, Howard H.;Li, Luowei;Lee, Maxwell P.;Huang, Jing;Yuspa, Stuart H.
氯细胞内通道 4 (CLIC4) 是 CLIC 家族中六种高度保守的蛋白质之一，与 GST 超家族中的 GST-omega 具有高度的结构同源性。虽然 CLIC4 是一种驻留在多个细胞区室中的多功能蛋白质，但其体外酶促谷氧还蛋白样活性的发现表明它可以充当抗氧化剂。在这里，我们发现使用 CRISPR 从小鼠 6DT1 乳腺肿瘤细胞中删除 CLIC4 会增强活性氧 (ROS) 的积累，并使细胞响应 H2O2 作为 ROS 诱导剂而对细胞凋亡敏感。在完整细胞中，H2O2 增加了 CLIC4 mRNA 和蛋白质的表达。此外，缺乏 CLIC4 的 6DT1 细胞中超氧化物产生的增加与线粒体过度活跃有关，包括线粒体膜电位增加和线粒体细胞器增大。然而，在缺乏 CLIC4 的情况下，H2O2 诱导的细胞凋亡与抗凋亡线粒体蛋白 Bcl2 和线粒体 ROS 负调节因子 UCP2 的低表达和降解有关。此外，H2O2 处理的对照细胞和 CLIC4-null 细胞的转录组分析显示，与 ROS 诱导的细胞凋亡相关的基因上调，以及维持线粒体功能的基因下调。因此，CLIC4缺陷型6DT1细胞移植形成的肿瘤高度坏死。这些结果强调了 CLIC4 在维持 6DT1 细胞中氧化还原稳态和线粒体功能中的关键作用。我们的研究结果还提出了靶向 CLIC4 来提高癌细胞对基于提高癌细胞中 ROS 的化疗药物的敏感性的可能性。
The chloride intracellular channel-4 (CLIC4) is one of the six highly conserved proteins in the CLIC family that share high structural homology with GST-omega in the GST superfamily. While CLIC4 is a multifunctional protein that resides in multiple cellular compartments, the discovery of its enzymatic glutaredoxin-like activity in vitro suggested that it could function as an antioxidant. Here, we found that deleting CLIC4 from murine 6DT1 breast tumor cells using CRISPR enhanced the accumulation of reactive oxygen species (ROS) and sensitized cells to apoptosis in response to H2O2 as a ROS-inducing agent. In intact cells, H2O2 increased the expression of both CLIC4 mRNA and protein. In addition, increased superoxide production in 6DT1 cells lacking CLIC4 was associated with mitochondrial hyperactivity including increased mitochondrial membrane potential and mitochondrial organelle enlargement. In the absence of CLIC4, however, H2O2-induced apoptosis was associated with low expression and degradation of the antiapoptotic mitochondrial protein Bcl2 and the negative regulator of mitochondrial ROS, UCP2. Furthermore, transcriptomic profiling of H2O2-treated control and CLIC4-null cells revealed upregulation of genes associated with ROS-induced apoptosis and downregulation of genes that sustain mitochondrial functions. Accordingly, tumors that formed from transplantation of CLIC4-deficient 6DT1 cells were highly necrotic. These results highlight a critical role for CLIC4 in maintaining redox-homeostasis and mitochondrial functions in 6DT1 cells. Our findings also raise the possibility of targeting CLIC4 to increase cancer cell sensitivity to chemotherapeutic drugs that are based on elevating ROS in cancer cells.
登录
查看更多内容
影响因子：
5.4
作者：
He, Guoan;Ma, Yao;Chou, Szu-Yi;Li, Huihong;Yang, Chingwen;Chuang, Jen-Zen;Sung, Ching-Hwa;Ding, Aihao
通讯作者：
Ding, Aihao
影响因子：
--
作者：
Gururaja Rao S;Ponnalagu D;Patel NJ;Singh H
通讯作者：
Singh H
影响因子：
4.8
作者：
Arnould, T;Mercy, L;Raes, M
通讯作者：
Raes, M
影响因子：
11.2
作者：
Alexandre, Jerome;Hu, Yumin;Huang, Peng
通讯作者：
Huang, Peng
低剂量tipifarnib对表达HIF-1α的胃癌细胞中MTOR信号通路和活性氧的产生的抗肿瘤作用。
影响因子：
2.6
作者：
Egawa N;Tanaka T;Matsufuji S;Yamada K;Ito K;Kitagawa H;Okuyama K;Kitajima Y;Noshiro H
通讯作者：
Noshiro H