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Effect of a poly(ADP-ribose) polymerase-1 inhibitor against esophageal squamous cell carcinoma cell lines.

Nasuno T, Mimaki S, Okamoto M, Esumi H, Tsuchihara K - Cancer Sci. (2013)

Bottom Line: Effective molecular target drugs that improve therapeutic efficacy with fewer adverse effects for esophageal cancer are highly anticipated.Of these eight cell lines, the clonogenic survival of one (TE-6) was reduced by AZD2281 to the level of DSB repair-defective Capan-1 and HCC1937 cells.Interestingly, a strong correlation between basal expression levels of γ-H2AX and sensitivity to AZD2281was observed in the TE-series cells (R(2)  = 0.5345).

View Article: PubMed Central - PubMed

Affiliation: Division of Translational Research, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Kashiwa, Chiba, Japan; Department of Otorhinolaryngology, University of Kitasato Hospital, Minami-ku, Sagamihara, Kanagawa, Japan.

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Increase in double strand breaks (DSBs) in TE-6 cells treated with AZD2281. (a) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and with 5 Gy X-ray irradiation, and γ-H2AX was assessed using Western blotting. The anti-γ-H2AX antibody detected both unubiquitinated (15 kD) and mono-ubiquitinated (23.6 kD) γ-H2AX. (b) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and γ-H2AX was assessed by immunofluorescence. DAPI (blue) and γ-H2AX (red) images were superimposed. (c) Number of the γ-H2AX-positive TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. (d) Scatter diagrams show the fluorescence intensity of individual TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. The lines shown indicated the averages of the data plotted. The data were obtained from at least 500 cells for each condition. **P < 0.01 (Student's t-test).
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fig03: Increase in double strand breaks (DSBs) in TE-6 cells treated with AZD2281. (a) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and with 5 Gy X-ray irradiation, and γ-H2AX was assessed using Western blotting. The anti-γ-H2AX antibody detected both unubiquitinated (15 kD) and mono-ubiquitinated (23.6 kD) γ-H2AX. (b) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and γ-H2AX was assessed by immunofluorescence. DAPI (blue) and γ-H2AX (red) images were superimposed. (c) Number of the γ-H2AX-positive TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. (d) Scatter diagrams show the fluorescence intensity of individual TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. The lines shown indicated the averages of the data plotted. The data were obtained from at least 500 cells for each condition. **P < 0.01 (Student's t-test).

Mentions: To determine whether DSBs are formed after treatment with AZD2281 for 24 h, we assessed the amount of γ-H2AX as a marker of DSBs. Western blotting revealed that the level of γ-H2AX staining in TE-6 cells increased in a dose-dependent manner. However, no such increase was observed in TE-1 cells (Fig. 3a). The same trend was observed by immunofluorescence. Both the percentage of γ-H2AX positive cells determined by visual inspection and the average fluorescence intensity of γ-H2AX staining per cell increased significantly in a dose-dependent manner in TE-6 cells, but not in TE-1 cells, suggesting that AZD2281 induced an accumulation of DNA damage in TE-6 cells (Fig. 3b–d).


Effect of a poly(ADP-ribose) polymerase-1 inhibitor against esophageal squamous cell carcinoma cell lines.

Nasuno T, Mimaki S, Okamoto M, Esumi H, Tsuchihara K - Cancer Sci. (2013)

Increase in double strand breaks (DSBs) in TE-6 cells treated with AZD2281. (a) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and with 5 Gy X-ray irradiation, and γ-H2AX was assessed using Western blotting. The anti-γ-H2AX antibody detected both unubiquitinated (15 kD) and mono-ubiquitinated (23.6 kD) γ-H2AX. (b) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and γ-H2AX was assessed by immunofluorescence. DAPI (blue) and γ-H2AX (red) images were superimposed. (c) Number of the γ-H2AX-positive TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. (d) Scatter diagrams show the fluorescence intensity of individual TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. The lines shown indicated the averages of the data plotted. The data were obtained from at least 500 cells for each condition. **P < 0.01 (Student's t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4317825&req=5

fig03: Increase in double strand breaks (DSBs) in TE-6 cells treated with AZD2281. (a) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and with 5 Gy X-ray irradiation, and γ-H2AX was assessed using Western blotting. The anti-γ-H2AX antibody detected both unubiquitinated (15 kD) and mono-ubiquitinated (23.6 kD) γ-H2AX. (b) TE-6 and TE-1 cells were treated with AZD2281 for 24 h and γ-H2AX was assessed by immunofluorescence. DAPI (blue) and γ-H2AX (red) images were superimposed. (c) Number of the γ-H2AX-positive TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. (d) Scatter diagrams show the fluorescence intensity of individual TE-6 and TE-1 cells treated with or without AZD2281 at the indicated concentrations for 24 h. The lines shown indicated the averages of the data plotted. The data were obtained from at least 500 cells for each condition. **P < 0.01 (Student's t-test).
Mentions: To determine whether DSBs are formed after treatment with AZD2281 for 24 h, we assessed the amount of γ-H2AX as a marker of DSBs. Western blotting revealed that the level of γ-H2AX staining in TE-6 cells increased in a dose-dependent manner. However, no such increase was observed in TE-1 cells (Fig. 3a). The same trend was observed by immunofluorescence. Both the percentage of γ-H2AX positive cells determined by visual inspection and the average fluorescence intensity of γ-H2AX staining per cell increased significantly in a dose-dependent manner in TE-6 cells, but not in TE-1 cells, suggesting that AZD2281 induced an accumulation of DNA damage in TE-6 cells (Fig. 3b–d).

Bottom Line: Effective molecular target drugs that improve therapeutic efficacy with fewer adverse effects for esophageal cancer are highly anticipated.Of these eight cell lines, the clonogenic survival of one (TE-6) was reduced by AZD2281 to the level of DSB repair-defective Capan-1 and HCC1937 cells.Interestingly, a strong correlation between basal expression levels of γ-H2AX and sensitivity to AZD2281was observed in the TE-series cells (R(2)  = 0.5345).

View Article: PubMed Central - PubMed

Affiliation: Division of Translational Research, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Kashiwa, Chiba, Japan; Department of Otorhinolaryngology, University of Kitasato Hospital, Minami-ku, Sagamihara, Kanagawa, Japan.

Show MeSH
Related in: MedlinePlus