Role Of NADPH Oxidase In X-ray Irradiation Induced HT-29, HeLa And GLC-82 Cell Damage

Objectives:â‘ Detect the change of cell survival after 2Gy X-ray irradiation which include HT-29 cell of low ray sensitive,HeLa cell of medium ray sensitive and GLC-82 cell of high ray sensitive.â‘¡Investigate the role of NADPH oxidase in X-ray irradiation induced HT-29,HeLa,GLC-82 cell damage.Methods:â‘ Choose good growth type of above three cells,divide them into control group and 2Gy X-ray irradiation group to detect the change of cell survival with MTT assay.â‘¡Divide them into control group,12Gy X-ray irradiation group, NADPH oxidase inhibitor group and NADPH oxidase inhibitor add 12Gy X-ray irradiation group,ROS was measured by fluorospectrophotometer.Immunostaining and confocal laser-scanning microscopy was employed to detect the co-localization of p47 and gp91phox,two subunit of NADPH oxidase.Western blotting were used to detect the expression of gp91phox before and after X-ray irradiation.Results:â‘ Cell survival decreased obviously after 2Gy X-ray irradiation.â‘¡Intra-cellular ROS increased obviously after 12Gy X-ray irradiation,but this increasing could be blocked by diphenyleneiodonium(DPI),an inhibitor of NADPH oxidase.Meanwhile,Cytosolic subunit p47phox movedto membrane and co-localizated with gp91phox.Western blotting indicated that gp91phox,the critical subunit of NADPH oxidase increased sharply after 12Gy X-ray irradiation.Conclusions:â‘ The survival HT-29 cell of low ray sensitive,HeLa cell of medium ray sensitive and GLC-82 cell of high ray sensitive were decreased after 2Gy X-ray irradiation.â‘¡NADPH oxidase mediated production of ROS play an important role in X-ray induced HT-29,HeLa,GLC-82 cell damage.