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Cited by 25Protein Kinase Regulation and GTPase SignalingEffects of Radiation ExposureCell death mechanisms and regulation

Ras-related small GTPases RalA and RalB regulate cellular survival after ionizing radiation

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Kidd, Ambrose R., Snider, Jared L., Martin, Timothy D., Graboski, Sarah F., Der, Channing J., Cox, Adrienne D.·2010·International Journal of Radiation Oncology*Biology*Physics
DOI 10.1016/j.ijrobp.2010.03.023PMC PMC2922474
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Abstract

Purpose Oncogenic activation of Ras renders cancer cells resistant to ionizing radiation (IR), but the mechanisms have not been fully characterized. The Ras-like small GTPases RalA and RalB are downstream effectors of Ras function and are critical for both tumor growth and survival. The Ral effector RalBP1/RLIP76 mediates survival of mice after whole-body irradiation, but the role of the Ral GTPases themselves in response to IR is unknown. We have investigated the role of RalA and RalB in cellular responses to IR. Methods and materials RalA, RalB, and their major effectors RalBP1 and Sec5 were knocked down by stable expression of short hairpin RNAs in the K-Ras-dependent pancreatic cancer-derived cell line MIA PaCa-2. Radiation responses were measured by standard clonogenic survival assays for reproductive survival, gammaH2AX expression for double-strand DNA breaks (DSBs), and poly(ADP-ribose)polymerase (PARP) cleavage for apoptosis. Results Knockdown of K-Ras, RalA, or RalB reduced colony-forming ability post-IR, and knockdown of either Ral isoform decreased the rate of DSB repair post-IR. However, knockdown of RalB, but not RalA, increased cell death. Surprisingly, neither RalBP1 nor Sec5 suppression affected colony formation post-IR. Conclusions Both RalA and RalB contribute to K-Ras-dependent IR resistance of MIA PaCa-2 cells. Sensitization due to suppressed Ral expression is likely due in part to decreased efficiency of DNA repair (RalA and RalB) and increased susceptibility to apoptosis (RalB). Ral-mediated radioresistance does not depend on either the RalBP1 or the exocyst complex, the two best-characterized Ral effectors, and instead may utilize an atypical or novel effector.

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Cited by

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Authors
Kidd, A. R., Snider, J. L., Martin, T. D., Graboski, S. F., Der, C. J., & Cox, A. D.
Affiliations
University of North Carolina at Chapel Hill (US)
Corresponding author
Cox, Adrienne D.
ORCID
Venue
International Journal of Radiation Oncology*Biology*Physics
Publisher
Elsevier BV
Published
2010-09
Volume
78
Issue
1
Pages
205-212
ISSN
0360-3016 (ISSN-L)
DOI
10.1016/j.ijrobp.2010.03.023
PMC
PMC2922474
OpenAlex
W2093879174
Cited by
25
References
0
Citations by year
2025: 1 · 2022: 3 · 2021: 1 · 2020: 2 · 2019: 1 · 2018: 2 · 2017: 1 · 2016: 4 · 2015: 5 · 2013: 1 · 2012: 2
FWCI
0.61
Type
journal article
Language
en
Open access
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • International Journal of Radiation Oncology*Biology*Physics (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • UNC Libraries
Funders
  • National Institutes of Health (5r01ca042978-30)
  • University of North Carolina at Chapel Hill
  • National Cancer Institute (CA042978, R01CA109550, CA 109550, R01-CA042978, U19CA067771, CA067771, F32CA138142)
  • National Institute of General Medical Sciences (T32GM007040)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
RalA, RalB, ionizing radiation resistance, pancreatic cancer, MIA PaCa-2 cells, DNA double-strand break repair, apoptosis, clonogenic survival, RALBP1, Sec5, exocyst complex, radiosensitization
MeSH terms
Cell Line, Cell Survival, Colony-Forming Units Assay, Humans, Pancreatic Neoplasms, Radiation Tolerance, Genes, ras, ATP-Binding Cassette Transporters, GTPase-Activating Proteins, ral GTP-Binding Proteins, Inverted Repeat Sequences, Gene Knockdown Techniques

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