In our previous studies, we showed that the apoptotic resistance of the human osteosarcoma cell line HS-Os-1 against irradiation was easily converted to a state of apoptotic-susceptibility by the addition of a relatively low concentration of hydrogen peroxide to the culture medium just prior to irradiation. When we consider the combined use of radiotherapy and hydrogen peroxide in a clinical setting for patients with radioresistant neoplasms, we need to be careful of the possible augmentation of the radiation effect to normal tissues of patients who undergo radiation therapy for their tumor in the presence of a low concentration of hydrogen peroxide in their topical tumor tissue. Therefore, we examined the combined effect of irradiation and hydrogen peroxide compared to that of irradiation alone for human peripheral T cells which were considered to be representative of normal tissue susceptible to apoptosis induced by irradiation. In this study, we compared the morphological changes in human peripheral T cells between both groups by utilizing MitoCapture, H2DCFDA (succinimidyl ester of dichloro-dihydrofluorescein diacetate), DAPI (4',6-diamidino-2-phenylindole), and LysoSensor. Our results showed that ROS formation was apparently augmented in the mitochondria and/or lysosomes instead of in the nuclei of irradiated T cells in the presence of a low concentration of hydrogen peroxide compared to those treated with irradiation alone. Moreover, dysfunction of mitochondrial membrane potential was also more evidently shown in human peripheral T cells irradiated under existence of a low concentration of hydrogen peroxide compared to T cells treated with 5 Gy irradiation alone. Based on these results, we concluded the possible existence of an augmentation effect of irradiation by the existence of a low concentration of hydrogen peroxide for human peripheral T cells. Therefore, we should be alert for the combined effects of radiation therapy and hydrogen peroxide on normal tissues in possible clinical situations when this combination is used for treatment of patients having radioresistant neoplasms such as osteosarcoma, malignant melanoma, and glioblastoma multiforme.

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