MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells.
MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells.
复制标题
MICAL1 通过调节乳腺癌细胞的氧化应激来控制细胞侵袭表型
DOI：
10.1186/s12885-016-2553-1
复制
发表时间：
2016-07-18
期刊：
影响因子：
3.8
通讯作者：
Du J
中科院分区：
文献类型：
--
作者：
Deng W;Wang Y;Gu L;Duan B;Cui J;Zhang Y;Chen Y;Sun S;Dong J;Du J
研究背景与CasL相互作用的分子（MICAL 1）是一种多结构域黄素蛋白单加氧酶，在促进癌细胞增殖和存活的机制中起重要作用。MICAL 1的激活导致HeLa细胞中活性氧（ROS）的上调。活性氧可以作为信号分子调节蛋白质磷酸化，导致癌细胞的恶性表型，例如侵袭和转移。在此，我们测试了MICAL 1是否可以控制细胞的迁移和侵袭，通过调节ROS在乳腺癌细胞line.MethodsThe消耗/过度表达的MICAL 1对细胞侵袭率的影响进行了测量，通过基于matrigel的transwell分析。采用CM 2-DCFHDA染色和增强光泽精化学发光法检测乳腺癌细胞内ROS含量。通过pulldown测定评估RAB 35活性。采用免疫荧光、免疫共沉淀、免疫印迹和共转染技术检测RAB 35和MICAL 1的相互关系。免疫印迹分析也被用来分析Akt磷酸化level.ResultsIn这项研究中，我们发现，MICAL 1的耗竭减少细胞迁移和侵袭以及ROS的产生。Akt的磷酸化也被MICAL 1耗竭减弱。同样，MICAL 1的过表达增加了ROS的产生，增加了Akt磷酸化，并有利于乳腺癌细胞的侵袭表型。此外，我们研究了EGF信号转导对MICAL 1功能的影响。我们证明了EGF增加了RAB 35的活化，并且活化形式的RAB 35可以与MICAL 1结合。沉默的RAB 35抑制ROS的产生，防止Akt磷酸化和抑制细胞侵袭响应EGF. ConclusionsTogether，我们的研究结果提供的证据表明，MICAL 1起着至关重要的作用，激活ROS/Akt信号和细胞侵袭表型，并确定一个新的RAB 35和MICAL 1之间的联系，在调节乳腺癌细胞的侵袭。这些发现可能为设计未来阻断乳腺癌转移的治疗策略提供依据。
BackgroundMolecules Interacting with CasL (MICAL1), a multidomain flavoprotein monoxygenase, is strongly involved in the mechanisms that promote cancer cell proliferation and survival. Activation of MICAL1 causes an up-regulation of reactive oxygen species (ROS) in HeLa cells. ROS can function as a signaling molecule that modulates protein phosphorylation, leading to malignant phenotypes of cancer cells such as invasion and metastasis. Herein, we tested whether MICAL1 could control cell migration and invasion through regulating ROS in breast cancer cell lines.MethodsThe effects of depletion/overexperssion of MICAL1 on cell invasion rate were measured by matrigel-based transwell assays. The contents of ROS in breast cancer cells were evaluated by CM2-DCFHDA staining and enhanced lucigenin chemiluminescence method. RAB35 activity was assessed by pulldown assay. The relationship of RAB35 and MICAL1 was evaluated by immunofluorescence, coimmunoprecipitation, immunoblotting and co-transfection techniques. Immunoblotting assays were also used to analyze Akt phosphorylation level.ResultsIn this study, we found that depletion of MICAL1 reduced cell migration and invasion as well as ROS generation. Phosphorylation of Akt was also attenuated by MICAL1 depletion. Likewise, the over-expression of MICAL1 augmented the generation of ROS, increased Akt phosphorylation, and favored invasive phenotype of breast cancer cells. Moreover, we investigated the effect of EGF signaling on MICAL1 function. We demonstrated that EGF increased RAB35 activation and activated form of RAB35 could bind to MICAL1. Silencing of RAB35 repressed ROS generation, prevented Akt phosphorylation and inhibited cell invasion in response to EGF.ConclusionsTaken together, our results provide evidence that MICAL1 plays an essential role in the activation of ROS/Akt signaling and cell invasive phenotype and identify a novel link between RAB35 and MICAL1 in regulating breast cancer cell invasion. These findings may provide a basis for designing future therapeutic strategy for blocking breast cancer metastasis.