Calpain 2 Cleavage of Akt in Glioblastoma Multiforme Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/m613n0455

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  • Glioblastoma is a malignant brain tumor that has extremely poor prognoses with a 5 year survival rate of less than 5%. The low survival rates of glioblastoma patients is attributed to glioblastoma cells being extremely resistant to radiotherapy and chemotherapy as well as being very invasive. We previously identified calpain 2, a calcium activated cysteine protease, to be required for invasion of glioblastoma. Here we identify a novel pathway in which calpain 2 directly cleaves Akt. Akt is a serine/threonine kinase that plays roles in numerous cellular processes including apoptosis, proliferation, and metabolism amongst others. We also identified through western blot analysis that calpain 2 is regulating cleavage of the isoforms Akt 1 and Akt 2. In addition we identified through immunofluorescence that calpain 2 appeared to be localized to the membrane ruffle region as well as the peri-nuclear region which is concomitant with Akt 2 and Akt 3 staining. We also identified that knockdown of calpain 2 in U87 glioblastoma cells caused a buildup of phosphorylated akt (ser473) as well as invadopodia at the membrane ruffle region indicating that calpain 2 may be regulating the turnover of invadopodia that are necessary for cell invasion.
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