Abstract Number: PB1801
Meeting: ISTH 2020 Congress
Background: Platelets can influence the phenotype of cancer cells through the release of eicosanoids. Among them, the 12-hydroxyeicosatetraenoic acid(12-HETE), a product of platelet-type 12-lipoxygenase(p12-LOX), is considered a key modulator in cancer metastasis.
Aims: We aimed to investigate the production of 12-HETE (both released and intracellular) in human platelets and colon cancer cells(HT-29) cultured alone, and whether their exposure to platelets influences 12-HETE generation and cancer cell phenotype via the p12-LOX pathway.
Methods: Human platelets(2×108) or HT-29(2×106) cells were cultured alone or cocultured, and the levels of 12-HETE evaluated by liquid chromatography-mass spectrometry(LC-MS/MS), gene expression by qPCR and p12-LOX protein by Western blot.
Results: In HT-29 cells, 12-HETE and p12-LOX were undetectable. In platelets, expressing the enzyme, 12-HETE was detected mainly intracellularly(0.418±0.138 nmol, mean±SD) versus the conditioned medium(0.018±0.004 nmol). Cancer cells exposed to platelets acquired the capacity to generate 12-HETE(extracellular levels: 0.013±0.004 nmol; intracellular levels: 0.024±0.007 nmol). In HT29 cells cultured with platelets, p12-LOX mRNA was undetectable while the protein was found. CDC(30µM), a selective inhibitor of p12-LOX, significantly reduced the levels of 12-HETE by 64%. In the coculture, platelet-derived microparticles(PMPs) were generated and expressed p12-LOX that was catalytically active. PMPs were internalized by HT29 cells, and this phenomenon was associated with the detection of p12-LOX in cancer cells and the capacity to synthesize 12-HETE. The crosstalk of platelets and cancer cells induced the expression of genes, such as ZEB1, vimentin, and fibronectin(involved in epithelial-mesenchymal transition), and RhoA(involved in cell migration). These effects were inhibited by the p12-LOX inhibitor CDC.
Conclusions: The interaction between platelets and HT29 cells caused the activation of platelets and the release of PMPs, which promoted the synthesis of 12-HETE via the transfer of p12-LOX to cancer cells. This phenomenon led to the induction of a more malignant phenotype of cancer cells, which was dampened by a p12-LOX inhibitor.
To cite this abstract in AMA style:Dovizio M, Schiavone S, Tacconelli S, Fullone R, Grande R, Zucchelli M, Marchisio M, Lanuti P, Ballerini P, Patrignani P. Generation of 12-HETE in Platelet-Cancer Cell Crosstalk [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/generation-of-12-hete-in-platelet-cancer-cell-crosstalk/. Accessed December 3, 2021.
« Back to ISTH 2020 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/generation-of-12-hete-in-platelet-cancer-cell-crosstalk/