Apoptosis induced by low‐dose and low‐dose‐rate radiation

BACKGROUND It has been claimed that external radiation, as a treatment modality for malignant diseases, partly induces apoptosis. It is not known, however, whether therapeutic low‐dose and low‐dose‐rate radiation are able to induce apoptosis. METHODS The effect of low‐dose radiation on apoptosis induction in HeLa Hep2 cells was studied, and quantitation of the apoptotic cells was performed by immunocytochemistry using TdT‐mediated dUtp‐x Nick End Labeling (TUNEL) technology and the M30 CytoDEATH antibody method. RESULTS When TUNEL staining was used to quantify apoptosis in untreated HeLa Hep2 cells kept in culture, approximately 5 ± 3% of the cells showed positive staining without any treatment. In the first experiment, the HeLa Hep2 cells were exposed to gamma radiation (i.e., 0.5, 1, 2, 5, 10, and 15 grays [Gy]) from a cobalt‐60 radiation source delivering a dose rate of 0.80 Gy/min. The radiated cells were cultivated for 5, 10, 24, 48, 72 and 168 hours after irradiation. Radiation doses below 2 Gy did not cause any significant apoptosis, but between 5 and 15 Gy significant apoptosis was observed, with peak values at 5 Gy (P < 0.001). Up to 60% of the investigated cells were shown to display apoptosis. Time to this peak value was 168 hours after irradiation. The HeLa Hep2 cells were exposed to doses of 2, 5, and 10 Gy at a 10‐fold lower dose rate (0.072 Gy/min). The cells that achieved a dose below 2 Gy did not present increased apoptosis. At doses above 2 Gy, however, the cells again demonstrated significant apoptosis. Up to 24 hours following irradiation, no apoptosis could be documented, whereas beyond 24 and up to 168 hours a highly significant apoptosis induction was observed. Significant cytotoxicity was confirmed by chromium‐51 release from the cells at 5 Gy. CONCLUSIONS Low‐dose and low‐dose‐rate radiation are able to induce significant apoptosis, and apoptosis may be one of the mechanisms by which low‐dose radiation causes growth inhibition. Cancer 2002;94:1210–4. © 2002 American Cancer Society. DOI 10.1002/cncr.10287 Low‐dose (2–5 grays) and low‐dose‐rate radiation (0.072 grays per minute) are able to induce significant apoptosis, and apoptosis may be one of the mechanisms by which low‐dose radiation causes growth inhibition of experimental tumors.