Cell Indu

نویسندگان

  • Ashley M. Young
  • Jean H. Overmeyer
  • William A. Maltese
چکیده

ownload huosis is a unique form of nonapoptotic cell death triggered by alterations in the trafficking of clathrinndent endosomes, ultimately leading to extreme vacuolization and rupture of the cell. Methuosis can be d in glioblastoma cells by expression of constitutively active Ras. This study identifies the small GTPases, nd Arf6, and the Arf6 GTPase-activating protein, GIT1, as key downstream components of the signaling ay underlying Ras-induced methuosis. The extent to which graded expression of active H-Ras(G12V) s cytoplasmic vacuolization correlates with the amount of endogenous Rac1 in the active GTP state. ng Rac1 activation with the specific Rac inhibitor, EHT 1864, or coexpression of dominant-negative 17N), prevents the accumulation of vacuoles induced by H-Ras(G12V). Coincident with Rac1 activa-Ras(G12V) causes a decrease in the amount of active Arf6, a GTPase that functions in the recycling hrin-independent endosomes. The effect of H-Ras(G12V) on Arf6 is blocked by EHT 1864, indicating e decrease in Arf6-GTP is directly linked to the activation of Rac1. Constitutively active Rac1(G12V) ts with GIT1 in immunoprecipitation assays. Ablation of GIT1 by short hairpin RNA prevents the se in active Arf6, inhibits vacuolization, and prevents loss of cell viability in cells expressing Rac1 ). Together, the results suggest that perturbations of endosome morphology associated with Ras-induced osis are due to downstream activation of Rac1 combined with reciprocal inactivation of Arf6. The seems to be mediated through Rac1 stimulation of GIT1. Further insights into this pathway could latter suggest opportunities for the induction of methuosis in cancers that are resistant to apoptotic cell death. Mol Cancer Res; 8(10); 1358–74. ©2010 AACR.

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تاریخ انتشار 2010