ABSTRACT
The aim of this study was to characterize genomic instability induced by ionizing radiation (IR) in human hepatocytes as reflected by alterations in cloning efficiency, micronucleus (MN) frequency, and apoptosis. The human normal liver 7702 cell line (HL7702) was subjected to initial irradiation of (60)Co-γ ray at doses of 0 (control group), 2, 4, 6, 8, or 10 Gy in each group. Progeny of surviving cells from a second irradiation at dose of 2 Gy were cultured for 15 passages until they were transferred. The cloning efficiency, MN frequency, and apoptotic rate were measured after the initial irradiation, and repeated at passage 15 before and after the second irradiation. The initial irradiation resulted in a dose-dependent decline in cloning efficiency and an increase in MN frequency and apoptotic rate. At passage 15 in progeny of initially irradiated cells, cloning efficiency, MN frequency returned to control levels while apoptotic rate rose. After the second irradiation, cloning efficiency fell while a rise in MN frequency and apoptosis occurred. Our results show that the second irradiation may further enhance cell progeny injury induced by initial irradiation, such that genomic instability that may be difficult to detect after one irradiation is more apparent with subsequent irradiation.
Subject(s)
Gamma Rays/adverse effects , Genomic Instability/radiation effects , Hepatocytes/radiation effects , Apoptosis/radiation effects , Cell Line , Cloning, Molecular/radiation effects , Dose-Response Relationship, Radiation , Humans , Micronuclei, Chromosome-Defective/radiation effects , Micronucleus TestsABSTRACT
This study was designed to characterize the differential protein expression in the progeny of human liver cells surviving exposure to ionizing radiation. The progeny of irradiated cells were derived from a human liver cell line exposed to 0, 2, 4, or 6 Gy of (60)Co gamma-irradiation. Total protein of the cells was extracted by two-dimensional electrophoresis (2-DE) and analyzed with ImageMaster 2D Platinum software. In total, 42 differentially expressed proteins from the progeny of irradiated cells were screened, of which 17 were identified by matrix assistant laser desorption ion-top flight-mass spectrometry (MALDI-TOF-MS) analysis. There were 4 upregulated and 13 downregulated proteins detected. The upregulated expression of two proteins, mitochondrial heat-shock 60-kD protein (HSP60) and globin transcription factor 1 (GATA-1), was further confirmed by immunoblotting. Database search revealed that these differentially expressed proteins may function in cell cycle regulation, cytoskeleton maintenance, stress response, and tumor metastasis, indicating an effect of radiation-induced genomic instability (RIGI) in the progeny of irradiated cells. Analysis on functional roles of the screened proteins may provide insight into further mechanistic investigations underlying molecular events induced by RIGI.
