TY - JOUR
T1 - RLIP76 in Defense of Radiation Poisoning
AU - Singhal, Jyotsana
AU - Singhal, Sharad S.
AU - Yadav, Sushma
AU - Suzuki, Sumihiro
AU - Warnke, Molly M.
AU - Yacoub, Adly
AU - Dent, Paul
AU - Bae, Sejong
AU - Sharma, Rajendra
AU - Awasthi, Yogesh C.
AU - Armstrong, Daniel W.
AU - Awasthi, Sanjay
N1 - Funding Information:
Supported in part by National Institutes of Health Grants CA 77495 and CA 104661 (to S.A.), Cancer Research Foundation of North Texas (to S.S. and S.Y.), Institute for Cancer Research, the Joe and Jessie Crump Fund for Medical Education (to S.S.), and the Robert A. Welch Foundation (Grant Y0026) (to M.M.W. and D.W.A.).
PY - 2008/10/1
Y1 - 2008/10/1
N2 - Purpose: To determine the role of RLIP76 in providing protection from radiation and chemotherapy. In the present report, we used RLIP76 to refer to both the mouse (Ralbp1) and the human (RLIP76) 76-kDa splice variant proteins (RLIP76) for convenience and to avoid confusion. In other reports, Ralbp1 refers to the mouse enzyme (encoded by the Ralbp1 gene), which is structurally and functionally homologous to RLIP76, the human protein encoded by the human RALBP1 gene. Methods and Materials: Median lethal dose studies were performed in RLIP76-/- and RLIP76+/+ C57B mice after treatment with a single dose of RLIP76 liposomes 14 h after whole body radiation. The radiosensitivity of the cultured mouse embryonic fibroblasts and the effects of buthionine sulfoximine (BSO), amifostine, c-jun N-terminal kinase (JNK), protein kinase B (Akt), and MAPK/ERK kinase (MEK) were determined by colony-forming assays. Glutathione-linked enzyme activities were measured by spectrophotometric assays, glutathione by dithiobis-2-nitrobenzoic acid (DTNB), lipid hydroperoxides by iodometric titration, and aldehydes and metabolites by thiobarbitauric acid reactive substances and liquid chromatography-mass spectrometry (LCMS). Results: RLIP76-/- mice were significantly more sensitive to radiation than were the wild-type, and RLIP76 liposomes prolonged survival in a dose-dependent manner in both genotypes. The levels of 4-hydroxynonenal and glutathione-conjugate of 4-hydroxynonenal were significantly increased in RLIP76-/- tissues compared with RLIP76+/+. RLIP76-/- mouse embryonic fibroblasts were markedly more radiosensitive than RLIP76+/+ mouse embryonic fibroblasts, despite increased glutathione levels in the former. RLIP76 augmentation had a remarkably greater protective effect compared with amifostine. The magnitude of effects of RLIP76 loss on radiation sensitivity was greater than those caused by perturbations of JNK, MEK, or Akt, and the effects of RLIP76 loss could not be completely compensated for by modulating the levels of these signaling proteins. Conclusion: The results of our study have shown that RLIP76 plays a central role in radiation resistance.
AB - Purpose: To determine the role of RLIP76 in providing protection from radiation and chemotherapy. In the present report, we used RLIP76 to refer to both the mouse (Ralbp1) and the human (RLIP76) 76-kDa splice variant proteins (RLIP76) for convenience and to avoid confusion. In other reports, Ralbp1 refers to the mouse enzyme (encoded by the Ralbp1 gene), which is structurally and functionally homologous to RLIP76, the human protein encoded by the human RALBP1 gene. Methods and Materials: Median lethal dose studies were performed in RLIP76-/- and RLIP76+/+ C57B mice after treatment with a single dose of RLIP76 liposomes 14 h after whole body radiation. The radiosensitivity of the cultured mouse embryonic fibroblasts and the effects of buthionine sulfoximine (BSO), amifostine, c-jun N-terminal kinase (JNK), protein kinase B (Akt), and MAPK/ERK kinase (MEK) were determined by colony-forming assays. Glutathione-linked enzyme activities were measured by spectrophotometric assays, glutathione by dithiobis-2-nitrobenzoic acid (DTNB), lipid hydroperoxides by iodometric titration, and aldehydes and metabolites by thiobarbitauric acid reactive substances and liquid chromatography-mass spectrometry (LCMS). Results: RLIP76-/- mice were significantly more sensitive to radiation than were the wild-type, and RLIP76 liposomes prolonged survival in a dose-dependent manner in both genotypes. The levels of 4-hydroxynonenal and glutathione-conjugate of 4-hydroxynonenal were significantly increased in RLIP76-/- tissues compared with RLIP76+/+. RLIP76-/- mouse embryonic fibroblasts were markedly more radiosensitive than RLIP76+/+ mouse embryonic fibroblasts, despite increased glutathione levels in the former. RLIP76 augmentation had a remarkably greater protective effect compared with amifostine. The magnitude of effects of RLIP76 loss on radiation sensitivity was greater than those caused by perturbations of JNK, MEK, or Akt, and the effects of RLIP76 loss could not be completely compensated for by modulating the levels of these signaling proteins. Conclusion: The results of our study have shown that RLIP76 plays a central role in radiation resistance.
KW - Embryonic fibroblasts
KW - RLIP76
KW - Radiation-resistance
KW - Ralbp1
UR - http://www.scopus.com/inward/record.url?scp=51449110220&partnerID=8YFLogxK
U2 - 10.1016/j.ijrobp.2008.06.1497
DO - 10.1016/j.ijrobp.2008.06.1497
M3 - Article
C2 - 18793957
AN - SCOPUS:51449110220
VL - 72
SP - 553
EP - 561
JO - International Journal of Radiation Oncology Biology Physics
JF - International Journal of Radiation Oncology Biology Physics
SN - 0360-3016
IS - 2
ER -