Active ras triggers death in glioblastoma cells through hyperstimulation of macropinocytosis.
Expression of activated Ras in glioblastoma cells induces accumulation of large phase-lucent cytoplasmic vacuoles, followed by cell death. This was previously described as autophagic cell death. However, unlike autophagosomes, the Ras-induced vacuoles are not bounded by a double membrane and do not sequester organelles or cytoplasm. Moreover, they are not acidic and do not contain the autophagosomal membrane protein LC3-II. Here we show that the vacuoles are enlarged macropinosomes. They rapidly incorporate extracellular fluid-phase tracers but do not sequester transferrin or the endosomal protein EEA1. Ultimately, the cells expressing activated Ras detach from the substratum and rupture, coincident with the displacement of cytoplasm with huge macropinosome-derived vacuoles. These changes are accompanied by caspase activation, but the broad-spectrum caspase inhibitor carbobenzoxy-Val-Ala-Asp-fluoromethylketone does not prevent cell death. Moreover, the majority of degenerating cells do not exhibit chromatin condensation typical of apoptosis. These observations provide evidence for a necrosis-like form of cell death initiated by dysregulation of macropinocytosis, which we have dubbed "methuosis." An activated form of the Rac1 GTPase induces a similar form of cell death, suggesting that Ras acts through Rac-dependent signaling pathways to hyperstimulate macropinocytosis in glioblastoma. Further study of these signaling pathways may lead to the identification of other chemical and physiologic triggers for this unusual form of cell death.
Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine, Block Health Sciences Building, Mail Stop 1010, 3035 Arlington Avenue, Toledo, OH 43614, USA.
Jean H
JH
Overmeyer
Aparna
A
Kaul
Erin E
EE
Johnson
William A
WA
Maltese
D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure*; D045744:Cell Line, Tumor; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure*; D006801:Humans; D009336:Necrosis; D010873:Pinocytosis; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism*; D014617:Vacuoles / Q000648:ultrastructure*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism
D016283:Proto-Oncogene Proteins p21(ras); D020830:rac1 GTP-Binding Protein