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The combination of ionizing radiation and expression of a wild type p53 gene via recombinant adenovirus induced a prominent tumour suppressing effect in human oral squamous cell carcinoma

Departments of ¹ Oral and Maxillofacial Radiology and ² Oral Microbiology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama City, Okayama 700-8525, JAPAN

View larger version (14K): [in a new window]   Figure 1. ß-galactosidase (ß-gal) activity in AxCAiLacZ-infected HSC4 and SAS cells. Cells were infected with AxCAiLacZ at multiplicity of infection (MOI) ranging from 0–25 plaque forming units per cell (pfu/cell). After 24 h, cells were lyzed and ß-galactosidase activity was determined. One unit of ß-galactosidase is defined as the amount of enzyme that will hydrolyze 1 µmol of ortho-nitrophenyl-ß-D-galactopyranoside in 1 min at 37°C.

View larger version (20K): [in a new window]   Figure 2. Detection of p53 production in the HSC4 and SAS cells by Western blot analysis using p53-specific monoclonal antibody. Cells were mock- or AxCAip53-infected and subsequently irradiated with 2 Gy. 24 h after irradiation, cell lysates were prepared. 50 µg of protein was loaded in each lane.

View larger version (19K): [in a new window]   Figure 3. Effects of AxCAip53 or AxCAiLacZ infection on radiation treatment of HSC4 and SAS cells. 24 h after infection, cells were irradiated with a single exposure of 0 Gy, 1 Gy, 2 Gy, 3 Gy, 4 Gy or 6 Gy. The number of colony forming units from non-infected and non-irradiated cells is defined as 100% of the surviving fraction. Each data point represents the mean value±standard deviation from three separate experiments. (a) HSC4 cells; (b) SAS cells. ——, mock-infected cells; ——, AxCAiLacZ-infected cells; ——, AxCAip53-infected cells.

View larger version (37K): [in a new window]   Figure 4. Flow cytometric analysis of HSC4 and SAS cells treated with AxCAip53 and irradiation. (a–d), HSC4 cells; (e–h), SAS cells. (a) and (e), mock-infected, non-irradiated cells; (b) and (f), mock-infected, irradiated cells; (c) and (g), AxCAip53-infected, non-irradiated cells; (d) and (h), AxCAip53-infected, irradiated cells.