Platelets promote invasion and induce epithelial to mesenchymal transition in ovarian cancer cells by TGF-β signaling pathway

Gynecol Oncol. 2019 Jun;153(3):639-650. doi: 10.1016/j.ygyno.2019.02.026. Epub 2019 Mar 28.

Abstract

Objective: To test whether platelets could increase invasion potential and initiate EMT in ovarian cancer cells via a TGF-β signaling pathway.

Methods: Blood samples were collected in 69 patients with ovarian cancer, 16 patients with benign ovarian tumor and 64 healthy donors. SK-OV-3 and OVCAR-3 ovarian cancer cells were treated with platelets. Transwell assays were used to analyze the invasive capacity, and EMT was assessed by microarray analysis, quantitative real-time PCR (qPCR) and Western blotting. Activation of TGF-β pathway was examined by ELISA and Western blotting. TGF-β type I receptor (TβR I) inhibitor A83-01 was used to confirm the role of TGF-β pathway in vitro and in vivo.

Results: Clinical data showed ovarian cancer patients with elevated platelet counts had a higher incidence of advanced stages. Treatment with platelets increased the invasive properties of both cell lines. Mesenchymal markers (snail family transcriptional repressor-1, vimentin, neural cadherin, fibronectin-1 and matrix metalloproteinase-2) were up-regulated in platelet-treated cells, while the epithelial marker (epithelial cadherin) was down-regulated. Higher TGF-β level was observed in patients with elevated platelet counts when compared to the subjects. Higher levels of TGF-β were also found in culture medium treated with platelets, and cells treated with platelets also showed increased phosphorylation of Smad2. TβR I inhibitor A83-01 reversed the EMT-like alterations and inhibited platelet-induced invasion in vitro and in vivo.

Conclusion: Platelet increased invasion potential and induced EMT in ovarian cancer cells in a TGF-β dependent pathway. Platelet-derived TGF-β may be useful as a new target treatment for ovarian cancer.

Keywords: A83–01; Epithelial to mesenchymal transition; Ovarian cancer; Platelet; TGF-β.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Blood Platelets*
  • Cadherins / genetics
  • Cadherins / metabolism
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Coculture Techniques
  • Epithelial-Mesenchymal Transition* / genetics
  • Female
  • Fibronectins / genetics
  • Gene Expression
  • Healthy Volunteers
  • Humans
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism
  • Mice
  • Middle Aged
  • Neoplasm Invasiveness
  • Neoplasm Transplantation
  • Ovarian Neoplasms / blood*
  • Ovarian Neoplasms / pathology
  • Platelet Count
  • Pyrazoles / pharmacology
  • Receptor, Transforming Growth Factor-beta Type I / antagonists & inhibitors
  • Signal Transduction
  • Smad Proteins / metabolism
  • Snail Family Transcription Factors / genetics
  • Thiosemicarbazones / pharmacology
  • Transforming Growth Factor beta / blood
  • Transforming Growth Factor beta / metabolism*
  • Up-Regulation
  • Vimentin / genetics

Substances

  • A-83-01
  • Cadherins
  • FN1 protein, human
  • Fibronectins
  • Pyrazoles
  • SNAI1 protein, human
  • Smad Proteins
  • Snail Family Transcription Factors
  • Thiosemicarbazones
  • Transforming Growth Factor beta
  • Vimentin
  • Receptor, Transforming Growth Factor-beta Type I
  • MMP2 protein, human
  • Matrix Metalloproteinase 2