Adenovirus-mediated wild-type p53 transfer radiosensitizes H1299 cells to subclinical-dose carbon-ion irradiation through the restoration of p53 function.
To determine whether adenovirus-mediated wild-type p53 transfer after radiotherapy could radiosensitize non-small-cell lung cancer (NSCLC) cells to subclinical-dose carbon-ion beam (C-beam), H1299 cells were exposed to a C-beam or gamma-ray and then infected with 5 MOI of AdCMV-p53 or GFP (C-beam or gamma-ray with p53 or GFP). Cell cycle was detected by flow cytometric analysis. The apoptosis was examined by a fluorescent microscope with DAPI staining. DNA fragmentation was monitored by the TUNEL assay. P53 mRNA was detected by reverse-transcriptase polymerase chain reaction. The expression of p53, MDM(2), and p21 was monitored by Western blot. Survival fractions were determined by colony-forming assay. The percentages of G(1)-phase cells in C-beam with p53 increased by 8.2%-16.0%, 5.2%-7.0%, and 5.8%-18.9%, respectively, compared with C-beam only, gamma-ray with p53, or p53 only. The accumulation of G(2)-phase cells in C-beam with p53 increased by 5.7%-8.9% and 8.8%-14.8%, compared with those in gamma-ray with p53 or p53 only, respectively. The percentage of apoptosis for C-beam with p53 increased by 7.4%-19.1%, 5.8%-11.7%, and 5.2 %-19.2%, respectively, compared with C-beam only, gamma-ray with p53, or p53 only. The level of p53 mRNA in C-beam with p53 was significantly higher than that in p53 only. The expression level of p53 and p21 in C-beam with p53 was significantly higher than that in both C-beam with GFP and p53 only. The survival fractions for C-beam with p53 were significantly less than those for the other groups (p < 0.05). The data suggested that AdCMV-p53 transfer could more efficiently radiosensitize H1299 cells to subclinical-dose C-beam irradiation through the restoration of p53 function.
Liu
Bing
B
Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Institute of Modern Physics, Chinese Academy of Sciences, 509 Nanchang Road, Lanzhou, China.
Zhang
Hong
H
Duan
Xin
X
Hao
Jifang
J
Xie
Yi
Y
Zhou
Qingming
Q
Wang
Yanling
Y
Tian
Yuan
Y
Wang
Tao
T
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D002244:Carbon / Q000737:chemistry; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000628:therapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D005434:Flow Cytometry; D016158:Genes, p53; D005822:Genetic Vectors; D006801:Humans; D007477:Ions; D008175:Lung Neoplasms / Q000235:genetics* / Q000628:therapy*; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D007477:Ions; D011838:Radiation-Sensitizing Agents; D016159:Tumor Suppressor Protein p53; D002244:Carbon