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 -ray and then infected with 5 MOI of AdCMV-p53 or GFP (C-beam or -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, MDM2, and p21 was monitored by Western blot. Survival fractions were determined by colony-forming assay. The percentages of G1-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, -ray with p53, or p53 only. The accumulation of G2-phase cells in C-beam with p53 increased by 5.7%–8.9% and 8.8%–14.8%, compared with those in -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, -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.

High-efficiency transfer and expression of AdCMV-p53 in human cervix adenocarcinoma cells induced by subclinical-dose carbon beam radiation

Purpose The aim of this study is to evaluate the eVect of carbon-beam irradiation on adenovirus-mediated p53 transfer in human cervix adenocarcinoma.Materials and methods The HeLa cells pre-exposed to carbon-beam or -ray, were infected with replication-deficient adenovirus recombinant vectors, containing human wild-type p53 (AdCMV-p53) and green Xuorescent protein (GFP) (AdCMV–GFP), respectively. The GFP transfer and p53 expression were detected by Xow cytometric analysis.Results The GFP transfer frequency in C-beam with AdCMV-GFP groups was 38–50% more than that inγ-ray with AdCMV–GFP groups. The percentage of p53 positive cells in the C-beam with AdCMV–p53 groups was 34–55.6% more than that in γ-ray with AdCMV-p53 groups (p < 0.05), suggesting that subclinical-dose C-beam irradiation could signiWcantly promote exogenous p53 transfer and p53 expression, and extend the duration of p53 expression in the HeLa cells. The expression of p21 increased with p53 expression in HeLa cells. The survival fractions for the 0.5–1.0 Gy C-beam with AdCMV-p53 groups were 38–43% less than those for the isodose γ-ray with AdCMV-p53 groups, and 31–40% less than those for the C-beam only groups (p <0.05).Conclusions The subclinical-dose C-beam irradiation could signiWcantly promote the transfer and expression of exogenous p53, extend the duration of p53 expression, and enhance the suppression of p53 on cervix adenocarcinoma cells.