Abstract
The present study examined the radiation biological response of cancer cells to different fractional irradiation doses and investigates the optimal fractional irradiation dose with improved biological effects. Radiobiological studies were performed at the molecular and cellular levels to provide insights into DNA damage and repair, and the apoptosis mechanism of cells that were exposed to different doses of X-ray irradiation (0, 2, 4, 6, 8, 10, 12.5, 15 and 20 Gy). Evidence of increased reactive oxygen species (ROS), DNA double strand breaks (DSB), cellular apoptosis, G2/M phase proportion and inhibition of cell proliferation were observed following irradiation. Differences in the ROS amount and apoptotic percentages of cells between the 2 and 4 Gy groups were insignificant. Compared with 0 Gy, the expression of the apoptosis suppression protein B-cell lymphoma-2 was decreased following at increased irradiation doses. However, apoptosis-associated protein Bcl-2-associated X (Bax), caspase-9 and BH3 interacting domain death agonist (Bid) were elevated following irradiation, compared with the control group (0 Gy). Furthermore, the expression levels of Bax in the 6, 8, 10 and 12.5 Gy groups were significantly increased, compared with the other groups. Caspase-9 expression with 2, 4, 6 and 8 Gy were increased compared with other groups, and the Bid levels with 6 and 8 Gy were also increased compared with other groups. G2/M phase arrest was associated with the increase of checkpoint kinase 1 and reduction of cyclin dependent kinase 1. DNA damage repair was associated with the protein Ku70 in the 2, 8, 10, 12.5, 15 and 20 Gy groups were less than other group. Compared with other group, Ku80 levels were reduced in the 6 and 8 Gy groups, and Rad51 levels were reduced in the 2, 8 and 10 Gy groups. The expression of hypoxia inducible factor-1α, c-Myc and glucose transporter 1 (GLUT1) demonstrated an increasing trend following irradiation in a dose-dependent manner, but the expression of pyruvate kinase M2, in the 2-10 Gy irradiation groups, and GLUT1, in the 12.5, 15 and 20 Gy irradiation groups, were reduced, compared with the other groups. Considering the DNA damage repair and apoptosis mechanisms at molecular and cellular levels, it was concluded that 2, 6, 8 and 10 Gy may be the optimal fractional dose that can promote cell apoptosis, and inhibit DNA damage repair and glycolysis.