title: E1A inhibition of radiation-induced NF-kappaB activity through suppression of IKK activity and IkappaB degradation, independent of Akt activation.
abstract: Activation of the transcription factor nuclear factor kappaB (NF-kappaB) has been implicated in the protection of cells from apoptosis. We have shown previously that the adenovirus type 5 E1A sensitizes cells to radiation-induced apoptosis by inhibiting NF-kappaB activity. However, the exact mechanism of inhibition is not known. In this study, we compared the activity of inhibitor of nuclear factor-kappaB (IkappaB) kinase (IKK) and the degradation of IkappaBalpha in E1A transfectants and parental human cancer cells after ionizing radiation treatment. We found that radiation-induced IKK activity and IkappaBalpha degradation were inhibited in the E1A transfectants. Recently, Akt has been implicated in NF-kappaB activation. To test whether Akt is regulated by E1A and is involved in radiation-induced NF-kappaB activity, we examined the phosphorylation status of Akt in the E1A transfectants and parental cells and in irradiated cells. The results indicated that radiation induced Akt phosphorylation and that E1A inhibited basal but not radiation-induced Akt phosphorylation. We additionally examined radiation-induced NF-kappaB activity in cells stably transfected with a dominant-negative, inactive Akt and in parental cancer cells treated with a phosphatidylinositol 3-kinase inhibitor, wortmannin. We found that dominant-negative Akt and wortmannin did not block radiation-induced NF-kappaB activity. Thus, our results suggest that inhibition of IKK activity and IkappaB degradation is the predominant mechanism for E1A-mediated inhibition of radiation-induced NF-kappaB activity and that radiation-induced Akt activation cannot be inhibited by E1A and is likely independent of radiation-induced NF-kappaB activity.
authors.affiliation: Department of Molecular and Cellular Oncology, Breast Cancer Basic Research Program, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
authors.forename: R
authors.initials: R
authors.lastname: Shao
authors.forename: E M
authors.initials: EM
authors.lastname: Tsai
authors.forename: K
authors.initials: K
authors.lastname: Wei
authors.forename: R
authors.initials: R
authors.lastname: von Lindern
authors.forename: Y H
authors.initials: YH
authors.lastname: Chen
authors.forename: K
authors.initials: K
authors.lastname: Makino
authors.forename: M C
authors.initials: MC
authors.lastname: Hung
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000502:physiology*; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D051550:I-kappa B Kinase; D020672:I-kappa B Proteins / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000528:radiation effects; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism / Q000532:radiotherapy; D010766:Phosphorylation / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D011836:Radiation Tolerance / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D017500:Adenovirus E1A Proteins; D020672:I-kappa B Proteins; D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; C494918:AKT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human