Enhanced radiosensitivity of malignant glioma cells after adenoviral p53 transduction.
OBJECTIVE
The goal of this study was to determine whether adenoviral vector-mediated expression of human wildtype p53 can enhance the radiosensitivity of malignant glioma cells that express native wild-type p53. The p53 gene is thought to function abnormally in the majority of malignant gliomas, although it has been demonstrated to be mutated in only approximately 30%. This has led to studies in which adenoviral transduction with wild-type human p53 has been investigated in an attempt to slow tumor cell growth. Recent studies suggest that reconstitution of wild-type p53 can render cells more susceptible to radiation-mediated death, primarily by p53-mediated apoptosis.


METHODS
Rat RT2 glioma cells were analyzed for native p53 status by reverse transcriptase-polymerase chain reaction and sequence analysis and for p53 expression by Western blot analysis. Clonogenic survival and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay were used to characterize RT2 cell radiosensitivity and apoptosis, respectively, with and without prior transduction with p53-containing and control adenoviral vectors. Animal survival length was monitored after intracerebral implantation with transduced and nontransduced RT2 cells, with and without cranial radiation. The RT2 cells were demonstrated to express native rat wild-type p53 and to markedly overexpress human p53 following adenoviral p53 transduction. The combination of p53 transduction followed by radiation resulted in marked decreases in RT2 cell survival and increases in apoptosis at radiation doses from 2 to 6 Gy. Animals receiving cranial radiation after intracerebral implantation with RT2 cells previously transduced with p53 survived significantly longer than control animals (p<0.01).


CONCLUSIONS
The ability to enhance the radiosensitivity of malignant glioma cells that express wild-type p53 by using adenoviral transduction to induce overexpression of p53 offers hope for this approach as a therapeutic strategy, not only in human gliomas that express mutant p53, but also in those that express wild-type p53.
Broaddus
W C
WC
Department of Radiation Oncology, Medical College of Virginia Hospital, Virginia Commonwealth University, Richmond, USA. wbroaddus@gems.vcu.edu
Liu
Y
Y
Steele
L L
LL
Gillies
G T
GT
Lin
P S
PS
Loudon
W G
WG
Valerie
K
K
Schmidt-Ullrich
R K
RK
Fillmore
H L
HL
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000528:radiation effects*; D001932:Brain Neoplasms / Q000235:genetics* / Q000532:radiotherapy; D002470:Cell Survival / Q000235:genetics / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D015316:Genetic Therapy; D005909:Glioblastoma / Q000235:genetics* / Q000532:radiotherapy; D005910:Glioma / Q000235:genetics* / Q000532:radiotherapy; D006801:Humans; D020287:In Situ Nick-End Labeling; D009368:Neoplasm Transplantation; D051381:Rats; D011916:Rats, Inbred F344; D014161:Transduction, Genetic; D014410:Tumor Stem Cell Assay; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
D016159:Tumor Suppressor Protein p53