Sonic hedgehog protects endometrial hyperplasial cells against oxidative stress via suppressing mitochondrial fission protein dynamin-like GTPase (Drp1)

Free Radic Biol Med. 2018 Dec:129:582-599. doi: 10.1016/j.freeradbiomed.2018.10.427. Epub 2018 Oct 19.

Abstract

Hh/Gli1 cascade as well as Gsk3β-Gli1 crosstalk play crucial role in estrogen-dependent progression of endometrial hyperplasia (EH). However, the underlying mechanisms involved in progression of disease still remain unclear. In the present study, we explored the role of Hh signaling in protection of endometrial hyperplasial cells against oxidative stress and the underlying mechanism involved therein. EH cells were found to be more resistant towards H2O2-induced oxidative stress (IC50: ~ 3×) as compared with normal endometrial cells. Estrogen (E2) pre-treatment followed by cytotoxic dose of H2O2, almost rescued the EH cells from apoptosis and caused the increased expression of downstream Shh signaling molecules i.e., Smo, Ptch and Gli1. Whereas pretreatment with cyclopamine was not able to curtail H2O2-induced effects indicating that estrogen protects these cells via activation of Shh pathway. Further, H2O2-induced ROS and lipid peroxidation alongwith decreased activities of antioxidant enzymes glutathione peroxidase and superoxide dismutase were found to be reversed in EH cells pre-exposed to E2 or rShh. The rShh suppressed H2O2-induced cell death and caused attenuation of mitochondrial apoptotic mediators and prevented disruption in mitochondrial morphology and mitochondrial membrane potential in EH cells. The functional blockage of signaling by Shh siRNA or Gli1siRNA led to significantly increased expression of mitochondrial fission protein dynamin-like GTPase (Drp1). The H2O2-treated EH cells showed diminished Gli1 and increased Drp1 expression, concurrent with reduced p-Drp1-(serine637). Whereas rShh pre-treated EH cells presented normal mitochondrial dynamics with dense, long networks of mitochondria alongwith nuclear accumulation of Gli1 and the decreased expression of Drp1. Overall, our results implicated that Shh signaling modulates antioxidant defense system and stabilizes mitochondrial dynamics by suppressing Drp1 protein which maintains survival of EH cells against oxidative stress.

Keywords: Dynamin-like GTPase; Endometrial hyperplasia; Estrogen; Mitochondrial dynamics; Shh signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Case-Control Studies
  • Disease Progression
  • Dynamins
  • Endometrial Hyperplasia / genetics*
  • Endometrial Hyperplasia / metabolism
  • Endometrial Hyperplasia / pathology
  • Endometrial Hyperplasia / surgery
  • Endometrium / metabolism
  • Endometrium / pathology
  • Endometrium / surgery
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • Estrogens / pharmacology
  • Female
  • GTP Phosphohydrolases / genetics*
  • GTP Phosphohydrolases / metabolism
  • Gene Expression Regulation
  • Glutathione Peroxidase / genetics
  • Glutathione Peroxidase / metabolism
  • Hedgehog Proteins / antagonists & inhibitors
  • Hedgehog Proteins / genetics*
  • Hedgehog Proteins / metabolism
  • Hedgehog Proteins / pharmacology
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Hysterectomy
  • Lipid Peroxidation / drug effects
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Mitochondrial Dynamics / drug effects*
  • Mitochondrial Proteins / genetics*
  • Mitochondrial Proteins / metabolism
  • Patched-1 Receptor / genetics
  • Patched-1 Receptor / metabolism
  • Primary Cell Culture
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Signal Transduction
  • Smoothened Receptor / genetics
  • Smoothened Receptor / metabolism
  • Superoxide Dismutase / genetics
  • Superoxide Dismutase / metabolism
  • Veratrum Alkaloids / pharmacology
  • Zinc Finger Protein GLI1 / antagonists & inhibitors
  • Zinc Finger Protein GLI1 / genetics*
  • Zinc Finger Protein GLI1 / metabolism

Substances

  • Estrogens
  • GLI1 protein, human
  • Hedgehog Proteins
  • Microtubule-Associated Proteins
  • Mitochondrial Proteins
  • PTCH1 protein, human
  • Patched-1 Receptor
  • RNA, Small Interfering
  • SHH protein, human
  • SMO protein, human
  • Smoothened Receptor
  • Veratrum Alkaloids
  • Zinc Finger Protein GLI1
  • Hydrogen Peroxide
  • Glutathione Peroxidase
  • Superoxide Dismutase
  • GTP Phosphohydrolases
  • DNM1L protein, human
  • Dynamins
  • cyclopamine