PGAM5-CypD pathway is involved in bromocriptine-induced RIP3/MLKL-dependent necroptosis of prolactinoma cells

Biomed Pharmacother. 2019 Mar:111:638-648. doi: 10.1016/j.biopha.2018.12.128. Epub 2019 Jan 3.

Abstract

Bromocriptine, the most commonly used dopamine (DA) receptor agonists for prolactinoma, can effectively reduce tumor size of prolactinoma, but the mechanism was not fully understood. Apoptosis had been well-recognized to contribute to the tumor mass regression caused by bromocriptine. However, whether other types of non-apoptotic cell death involved in the bromocriptine-induced prolactinoma shrinkage had not been fully clarified. The newly discovered molecular mechanism of necroptosis provides the possibility to examine this programmed necrosis in the pharmacological function of bromocriptine. The aim of present study was to evaluate and investigate the underlying mechanism of necroptosis in involution of prolactinoma induced by bromocriptine. By immunohistochemistry, we found that the numbers of receptor-interacting serine-threonine kinase 3(RIP3) and phosphorylated mixed lineage kinase domain-like protein (pMLKL)-positive cells and their expression intensities were increased in patients with prolactinoma after bromocriptine therapy. For further exploring the mechanism of bromocriptine, prolactinoma cell line (MMQ cells) was adopted to study the mechanism of necroptosis in vitro. Cell viability and ATP level of MMQ cells were decreased, while reactive oxygen species (ROS) level was increased after bromocriptine treatment. The above effects could be partially reversed by Necrostatin-1, an inhibitor of necroptosis. Ultrastructural study further confirmed the necroptosis of MMQ cells, which was characterized by ruptured membrane, dissolved cytoplasm and especially the dramatically swollen mitochondria. Furthermore, we demonstrated that bromocriptine induced RIP3/MLKL-dependent necroptosis of prolactinoma cells and phosphoglycerate mutase family 5(PGAM5)/ Cyclophilin D (CypD) pathway was involved. The results suggested that necroptosis might be a promising target for clinical therapy for prolactinoma.

Keywords: Bromocriptine; MMQ cells; Necroptosis; Prolactinoma.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Bromocriptine / pharmacology*
  • Bromocriptine / therapeutic use
  • Cell Survival
  • Cyclophilins / metabolism*
  • Dopamine Agonists / pharmacology
  • Dopamine Agonists / therapeutic use
  • Dose-Response Relationship, Drug
  • Female
  • Humans
  • Male
  • Middle Aged
  • Mitochondrial Proteins / metabolism*
  • Necrosis / drug therapy
  • Necrosis / metabolism
  • Necrosis / pathology
  • Phosphoprotein Phosphatases / metabolism*
  • Prolactinoma / drug therapy
  • Prolactinoma / metabolism*
  • Prolactinoma / pathology
  • Protein Kinases / metabolism*
  • Rats
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Young Adult

Substances

  • Dopamine Agonists
  • Mitochondrial Proteins
  • Bromocriptine
  • MLKL protein, human
  • Protein Kinases
  • RIPK3 protein, human
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • PGAM5 protein, human
  • Phosphoprotein Phosphatases
  • Cyclophilins
  • PPID protein, human