Activation of cyclic AMP/PKA pathway inhibits bladder cancer cell invasion by targeting MAP4-dependent microtubule dynamics

Urol Oncol. 2014 Jan;32(1):47.e21-8. doi: 10.1016/j.urolonc.2013.06.017. Epub 2013 Oct 17.

Abstract

Objective: With the notorious reputation of the vicious invasion, the bladder cancer is the most common malignant tumor of the urinary system. Inhibiting invasion through microtubule dynamics interruption has emerged as an important treatment of bladder cancer. Here we investigated the role of the cyclic adenosine monophosphate/protein kinase A (cAMP/PKA) pathway in human bladder cancer cells invasion.

Materials and methods: With or without the treatment of various cAMP elevators, we assessed invasive and migrated capabilities of T24 and UM-UC-3, two high-grade invasive bladder cancer cell lines, using matrigel transwell inserts assay and scratch wound healing assay. The microtubule (MT) dynamics were examined by immunofluorescence and immunoblotting. Microtubule-Associated Protein 4 (MAP4) was silenced to investigate its role in tumor invasion. We also analyzed gene expression of MAP4 in 34 patients with bladder cancer using immunohistochemical staining assay. The interaction between PKA and MAP4 was examined by co-immunoprecipitation.

Results: We used cAMP elevators and small interfering RNA of MAP4 here, found that both of them can potently inhibit the invasion and the migration of bladder cancer cells by disrupting microtubule (MT) cytoskeleton. Consistently, the bladder cancer grade is positively correlated with the protein level of MAP4. Furthermore, we found that cAMP/PKA signaling can disrupt MT cytoskeleton by the phosphorylation of MAP4.

Conclusion: Our results indicated that the cAMP/PKA signaling pathway might inhibit bladder cancer cell invasion by targeting MAP4-dependent microtubule dynamics, which could be exploited for the therapy of invasive bladder cancer.

Keywords: Cyclic AMP/PKA pathway; Human bladder cancer; Invasion; MAP4; Microtubules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bucladesine / pharmacology
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • Cell Movement / physiology*
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Cholera Toxin / pharmacology
  • Cyclic AMP / metabolism*
  • Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / genetics
  • Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / metabolism*
  • Cytoskeleton / drug effects
  • Cytoskeleton / metabolism
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Microscopy, Confocal
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / drug effects
  • Microtubules / metabolism
  • Neoplasm Invasiveness
  • Phosphorylation / drug effects
  • Protein Binding
  • RNA Interference
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Signal Transduction / physiology*
  • Tubulin / metabolism
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / metabolism*
  • Urinary Bladder Neoplasms / pathology

Substances

  • Microtubule-Associated Proteins
  • Tubulin
  • Bucladesine
  • Cholera Toxin
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinase Catalytic Subunits