Suppression subtractive hybridization in construction of radiation-induced EST library

Objective To clone and identify radiation\|induced genes from 0 5 Gy γ ray irradiated human embryo lung cells(HEL).The identification and functional studies of radiation\|induced genes will prompt the elucidation of the molecular mechanisms of low dose radiation\|induced biological effects.It will also profit the understanding of the basic processes of cellular metabolisms. Methods A low dose radiation\|induced differentially displaying EST library has been constructed by suppression subtractive hybridization from HEL cells irradiated with 0 5 Gy γ rays.The EST library was screened by reverse Northern hybridization analysis.Positive clones were sequenced and the similarity was searched against the DNA database in GenBank. Results Altogether 90 positive cDNA clones with increased expression after 0 5 Gy irradiation were identified which corresponded to 21 individual genes.These genes involved in the processes of cell cycle control,signal transduction,cell skeleton,metabolism and protein synthesis,etc.All these demonstrated the diverse responses of cells to low dose radiation.Moreover,four novel cDNA were obtained. Conclusions Low dose radiation induces ESTs which relate to cell proliferation,cell cycle control,signal transduction,cell skeleton,metabolism,protein synthesis and stress response etc were cloned and identified by SSH.Our data suggest that these genes could play important roles in the biological response of cells to low dose radiation.\;