title: Radiation enhances the invasive potential of primary glioblastoma cells via activation of the Rho signaling pathway.
abstract: Glioblastoma multiforme (GBM) is among the most treatment-refractory of all human tumors. Radiation is effective at prolonging survival of GBM patients; however, the vast majority of GBM patients demonstrate progression at or near the site of original treatment. We have identified primary GBM cell lines that demonstrate increased invasive potential upon radiation exposure. As this represents a novel mechanism by which radiation-treated GBMs can fail therapy, we further investigated the identity of downstream signaling molecules that enhance the invasive phenotype of irradiated GBMs. Matrigel matrices were used to compare the extent of invasion of irradiated vs. non-irradiated GBM cell lines UN3 and GM2. The in vitro invasive potential of these irradiated cells were characterized in the presence of both pharmacologic and dominant negative inhibitors of extracellular matrix and cell signaling molecules including MMP, uPA, IGFR, EGFR, PI-3K, AKT, and Rho kinase. The effect of radiation on the expression of these signaling molecules was determined with Western blot assays. Ultimately, the in vitro tumor invasion results were confirmed using an in vivo 9L GBM model in rats. Using the primary GBM cell lines UN3 and GM2, we found that radiation enhances the invasive potential of these cells via activation of EGFR and IGFR1. Our findings suggest that activation of Rho signaling via PI-3K is required for radiation-induced invasion, although not required for invasion under physiologic conditions. This report clearly demonstrates that radiation-mediated invasion is fundamentally distinct from invasion under normal cellular physiology and identifies potential therapeutic targets to overcome this phenomenon.
authors.affiliation: Department of Radiation Oncology, Massachusetts General Hospital/Harvard Medical School, Boston, MA, 02114, USA.
authors.forename: Gary G
authors.initials: GG
authors.lastname: Zhai
authors.forename: Rajeev
authors.initials: R
authors.lastname: Malhotra
authors.forename: Meaghan
authors.initials: M
authors.lastname: Delaney
authors.forename: Douglas
authors.initials: D
authors.lastname: Latham
authors.forename: Ulf
authors.initials: U
authors.lastname: Nestler
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Neelanjan
authors.initials: N
authors.lastname: Mukherjee
authors.forename: Qinhui
authors.initials: Q
authors.lastname: Song
authors.forename: Pierre
authors.initials: P
authors.lastname: Robe
authors.forename: Arnab
authors.initials: A
authors.lastname: Chakravarti
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D004789:Enzyme Activation / Q000528:radiation effects; D066246:ErbB Receptors / Q000378:metabolism / Q000528:radiation effects; D005909:Glioblastoma / Q000473:pathology*; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism / Q000528:radiation effects; D011836:Radiation Tolerance / Q000502:physiology*; D051381:Rats; D011916:Rats, Inbred F344; D017526:Receptor, IGF Type 1 / Q000378:metabolism / Q000528:radiation effects; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000378:metabolism / Q000528:radiation effects*
chemical_list: D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D017526:Receptor, IGF Type 1; D020741:rho GTP-Binding Proteins