title: MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells.
abstract: Molecules 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.
  
  
  
  The 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.
  
  
  
  In 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.
  
  
  
  Taken 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.
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.|Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Deng
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Yueyuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.|Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China.|Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Luo
authors.initials: L
authors.lastname: Gu
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Duan
authors.affiliation: Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China.|Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Cui
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.|Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China.|Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China.|Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Shixiu
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 211166, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Dong
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, 211166, China. dujun@njmu.edu.cn.|Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, 211166, China. dujun@njmu.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Du
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D049109:Cell Proliferation; D002470:Cell Survival; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D060588:LIM Domain Proteins / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins; D006899:Mixed Function Oxygenases; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology*; D018384:Oxidative Stress / Q000502:physiology; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000096985:Calponins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; D060588:LIM Domain Proteins; D008840:Microfilament Proteins; D006899:Mixed Function Oxygenases; D051057:Proto-Oncogene Proteins c-akt; D017382:Reactive Oxygen Species; D020691:rab GTP-Binding Proteins; C455272:MICAL1 protein, human; D000096985:Calponins; C511183:RAB35 protein, human
keywords: Breast cancer; EGF; Invasion; MICAL1; RAB35; ROS