Effects of FTO gene on the radiosensitivity of A172 cells and its mechanism / 中华放射医学与防护杂志

ABSTRACT

Objective To evaluate the effect of Fat Mass and Obesity Associated (FTO) gene on radiosensitivity of human glioma cell A172 and investigate its potential mechanism by changing the expression of FTO gene.Methods Cells were divided into five groups according to their FTO protein expression level.The normal expression group was recorded as A172 Group,the low-expression and its negative control group was A172/siRNA and A172/NC Group,and the over-expression and its negative control group was A172/FTO and A172/PC group.FTO protein expressions were assayed by Western blot in A172 Group after irradiation.Clonogenic assay was executed to evaluate the relationship between FTO gene and radiosensitivity.Immunofluorescence and Western blot assay were applied to detect the proteins of DNA damage and repair.Results FTO protein expression level in A172 Group was significantly related to the irradiation dose and the time post-irradiation.The radiosensitization ratio (SERD0) of A172/siRNA and A172/FTO group were 1.829 and 0.812 respectively.Not only the number of γ-H2AX foci increased (t =-21.884,P ＜ 0.05) in A172/siRNA 1 h post-irradiation but the decreases of p-p95/NBS1 and Ku80 proteins were also detected (t =24.731,23.293,P ＜ 0.05) together with the increase of Rad50 protein (t =3.140,P ＜ 0.05).But the expressions of these proteins in A172/FTO group were opposite to the above phenomenon (t =0.642,-8.364,26.829,P ＜ 0.05).Conclusions FTO gene is a radiation-resistant gene,which may because the regulation of FTO gene could alter the primary injury and DNA damage repair in the irradiated tumor cells.