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Benzidine Induces Epithelial-Mesenchymal Transition of Human Bladder Cancer Cells through Activation of ERK5 Pathway

  • Sun, Xin (Department of Urology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Zhang, Tao (Department of Urology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Deng, Qifei (Second Department of Urology, Anhui Provincial Children's Hospital) ;
  • Zhou, Qirui (Department of Otolaryngology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Sun, Xianchao (Department of Urology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Li, Enlai (Department of Urology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Yu, Dexin (Department of Urology, The Second Affiliated Hospital of Anhui Medical University) ;
  • Zhong, Caiyun (Department of Nutrition and Food Safety, School of Public Health, Nanjing Medical University)
  • Received : 2017.07.02
  • Accepted : 2017.12.20
  • Published : 2018.03.31

Abstract

Benzidine, a known carcinogen, is closely associated with the development of bladder cancer (BC). Epithelial-mesenchymal transition (EMT) is a critical pathophysiological process in BC progression. The underlying molecular mechanisms of mitogen-activated protein kinase (MAPK) pathway, especially extracellular regulated protein kinases 5 (ERK5), in regulating benzidine-induced EMT remains unclarified. Hence, two human bladder cell lines, T24 and EJ, were utilized in our study. Briefly, cell migration was assessed by wound healing assay, and cell invasion was determined by Transwell assay. Quantitative PCR and western blot were utilized to determine both gene expressions as well as protein levels of EMT and MAPK, respectively. Small interfering RNA (siRNA) was transfected to further determine ERK5 function. As a result, the migration and invasion abilities were enhanced, epithelial marker expression was decreased while mesenchymal marker expression was increased in human BC cell lines. Meanwhile, benzidine administration led to activation of ERK5 and activator protein 1 (AP-1) proteins, without effective stimulation of the Jun N-terminal kinase (JNK) or p38 pathways. Moreover, Benzidine-induced EMT and ERK5 activation were completely suppressed by XMD8-92 and siRNAs specific to ERK5. Of note, ERK1/2 was activated in benzidine-treated T24 cells, while benzidine-induced EMT could not be reversed by U0126, an ERK1/2 inhibitor, as indicated by further study. Collectively, our findings revealed that ERK5-mediated EMT was critically involved in benzidine-correlated BC progression, indicating the therapeutic significance of ERK5 in benzidine-related BC.

Acknowledgement

Supported by : National Natural Science Foundation of China

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