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Cell J. 2011 Summer;13(2):83-90. Epub 2011 Aug 24.

Evaluation of the Combined Effect of 2ME2 and (60)Co on the Inducement of DNA Damage by IUdR in a Spheroid Model of the U87MG Glioblastoma Cancer Cell Line Using Alkaline Comet Assay.

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1. Medical Physics Department, School of Medical Basic Sciences, Tehran University of Medical Sciences, Tehran, Iran ; 2. Cellular and Molecular Research Center, Tehran University of Medical Sciences, Tehran, Iran.



In this study, we investigated the combined effect of 2-Methoxyestradiol (2ME2) and (60)Co on the cytogenetic damage of iododeoxyuridine (IUdR) in the spheroid model of U87MG glioblastoma cancer cell lines by alkaline comet assay.


U87MG cells were cultured as spheroids with diameters of 350 µm. As control, the spheroids of one plate were not treated. Other cultures were pretreated with 2ME2 (250 µM) for one volume doubling time (1 VDT). After this time, the subsequent treatments were performed according to the following groups: Vehicle (this sample was not treated in the 2(nd) VDT) Treated with 2ME2 (250 µM) for 1 VDT Treated simultaneously with 2ME2 (250 µM) and IUdR (1 µM) for 1 VDT Treated with 2ME2 (250 µM) for 1 VDT then irradiated with (60)Co (2 Gy) Treated simultaneously with 2ME2 (250 µM) and IUdR (1 µM) for 1 VDT then irradiated with (60)Co (2 Gy) Then the DNA damage was evaluated using the alkaline comet assay method.


The results showed that 2ME2 in combination with gamma irradiation of (60)Co significantly (p<0.001) increased the DNA damage by IUdR as compared to the control group. Thus the combination of these two agents increased the cytogenetic effects of IUdR in the spheroid culture model of U87MG glioblastoma cell lines.


By inhibiting the HIF-1α protein and preventing the G0 phase arrest, 2ME2 causes an increased progression into S phase and increases the IUdR absorption. Then the DNA damage in the spheroid cells increases as the uptake of IUdR is increased. These results suggest that the combined use of 2ME2 and (60)Co can increase the radiosensitization effect of IUdR.


2-Methoxyestradiol; Comet; DNA Damage; HIF-1Alpha; Iododeoxyuridinre

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