Diarachidonoylphosphoethanolamine induces necrosis/necroptosis of malignant pleural mesothelioma cells

Our plain-English summary of Diarachidonoylphosphoethanolamine induces necrosis/necroptosis of malignant pleural mesothelioma cells, by Y. Kaku, A. Tsuchiya, T. Kanno, et al (Cellular Signalling, 2015).

The study

• Authors: Y. Kaku, A. Tsuchiya, T. Kanno, T. Nakano, T. Nishizaki
• Journal: Cellular Signalling
• Published: 2015;27(9):1713-9
• Cited by: 8 articles, as counted by NIH iCite
• Read it: PubMed · Publisher

Abstract

The present study investigated 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine (DAPE)-induced cell death in malignant pleural mesothelioma (MPM) cells. DAPE reduced cell viability in NCI-H28, NCI-H2052, NCI-H2452, and MSTO-211H MPM cell lines in a concentration (1-100μM)-dependent manner. In the flow cytometry using propidium iodide (PI) and annexin V (AV), DAPE significantly increased the population of PI-positive and AV-negative cells, corresponding to primary necrosis, and that of PI-positive and AV-positive cells, corresponding to late apoptosis/secondary necrosis, in NCI-H28 cells. DAPE-induced reduction of NCI-H28 cell viability was partially inhibited by necrostatin-1, an inhibitor of RIP1 kinase to induce necroptosis, or knocking-down RIP1. DAPE generated reactive oxygen species (ROS) followed by disruption of mitochondrial membrane potentials in NCI-H28 cells. DAPE-induced mitochondrial damage was attenuated by cyclosporin A, an inhibitor of cyclophilin D (CypD). DAPE did not affect expression and mitochondrial localization of p53 protein in NCI-H28 cells. DAPE significantly decreased intracellular ATP concentrations in NCI-H28 cells. Overall, the results of the present study indicate that DAPE induces necroptosis and necrosis of MPM cells; the former is mediated by RIP1 kinase and the latter is caused by generating ROS and opening CypD-dependent mitochondrial permeability transition pore, to reduce intracellular ATP concentrations.

Copyright © 2015 Elsevier Inc. All rights reserved.

In plain English

This was a laboratory study of cancer cells grown in dishes. It involved no patients and no asbestos exposure. The authors tested a fat-like molecule called DAPE on four malignant pleural mesothelioma cell lines: NCI-H28, NCI-H2052, NCI-H2452 and MSTO-211H. After 24 hours, DAPE reduced measured cell viability (an MTT test signal) in all four lines, more so at higher doses from 1 to 100 micromolar, reaching nearly 0% of untreated levels at 100 micromolar. Most later tests used only NCI-H28 cells. In them, DAPE increased cells showing necrosis (uncontrolled cell death) and late apoptosis or secondary necrosis, and halted cell division at certain stages. With 100 micromolar DAPE, a RIP1-blocking drug partly prevented the loss of viability. Lowering the amount of RIP1 protein with a gene-silencing tool (siRNA) reduced cell death at that dose but not at 10 to 70 micromolar. The authors read this as a sign of necroptosis, a programmed form of necrosis. DAPE also raised reactive oxygen species, disrupted mitochondria, and lowered cell energy (ATP). It did not change p53 protein levels. The authors concluded that DAPE causes necroptosis and necrosis of these mesothelioma cells.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory experiment on cancer cell lines grown in culture (in vitro)
• Who was studied: Four malignant pleural mesothelioma cell lines (NCI-H28, NCI-H2052, NCI-H2452, MSTO-211H), bought from the American Type Culture Collection. Most mechanism experiments used NCI-H28 only. Most experiments were repeated 4 times, and 8 times for NCI-H2452 in the viability test. No patients or animals were studied.
• Exposure: No asbestos exposure was studied. Cells were treated with DAPE (1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine) at 1 to 100 micromolar for 24 hours in viability tests, at 30 micromolar for 24 or 48 hours in flow cytometry, and at 100 micromolar in most mechanism experiments.
• Main results:
  • After 24 hours, DAPE reduced cell viability, measured by MTT assay signal relative to untreated cells, in all four mesothelioma cell lines. The reduction grew with concentration (1 to 100 micromolar) and reached nearly 0% of basal levels at 100 micromolar.
  • In NCI-H28 cells, DAPE significantly increased the share of cells classed as primary necrosis (PI-positive, AV-negative). It also increased the share classed as late apoptosis or secondary necrosis (PI-positive, AV-positive).
  • In NCI-H28 cells, DAPE increased the proportion of cells in the S and G2/M phases and decreased the proportion in G1. The authors interpreted this as cell cycle arrest.
  • In NCI-H28 cells treated with 100 micromolar DAPE, necrostatin-1 (10 micromolar), a RIP1 kinase inhibitor, partially prevented the loss of viability.
  • In NCI-H28 cells, knocking down RIP1 with siRNA significantly suppressed cell death caused by 100 micromolar DAPE. It did not affect cell death at lower concentrations (10 to 70 micromolar).
  • In NCI-H28 cells, DAPE raised reactive oxygen species starting at 1 minute, with a peak at 10 minutes. The scavenger N-acetyl-L-cysteine abolished the rise.
  • In NCI-H28 cells, DAPE disrupted mitochondrial membrane potentials starting at 3 minutes. Cyclosporin A, an inhibitor of cyclophilin D, prevented this damage.
  • In NCI-H28 cells, DAPE did not affect basal p53 protein levels and did not increase the amount of p53 in mitochondria.
  • In NCI-H28 cells, DAPE significantly reduced intracellular ATP concentrations compared with before treatment.
• The authors' conclusion: The authors concluded that DAPE induces RIP1 kinase-mediated necroptosis of malignant pleural mesothelioma cells. They concluded that it also, alternatively, induces necrosis: reactive oxygen species are produced, cyclophilin D-dependent mitochondrial permeability transition follows, and intracellular ATP falls.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Necrosis, Pleural Mesothelioma, Apoptosis, Reactive Oxygen Species (ROS).

Related studies

• Diarachidonoylphosphoethanolamine induces apoptosis of malignant pleural mesothelioma cells through a Trx/ASK1/p38 MAPK pathway — A. Tsuchiya, Y. Kaku, et al. (2015)
• Pifithrin-μ induces necroptosis through oxidative mitochondrial damage but accompanies epithelial-mesenchymal transition-like phenomenon in malignant mesothelioma cells under lactic acidosis — Y.J. Lee, K.S. Park, et al. (2019)
• Particles of different sizes and shapes induce neutrophil necroptosis followed by the release of neutrophil extracellular trap-like chromatin — J. Desai, O. Foresto-Neto, et al. (2017)
• Phosphatidylinositol Induces Caspase-Independent Apoptosis of Malignant Pleural Mesothelioma Cells by Accumulating AIF in the Nucleus — S. Kanemura, A. Tsuchiya, et al. (2015)
• AMP converted from intracellularly transported adenosine upregulates p53 expression to induce malignant pleural mesothelioma cell apoptosis — Y. Nogi, T. Kanno, et al. (2012)
• Programmed necrosis induced by asbestos in human mesothelial cells causes high-mobility group box 1 protein release and resultant inflammation — H. Yang, Z. Rivera, et al. (2010)

Browse return to medical research.