Project description:In order to identified as candidate biomarkers for early diagonosis or as therapeutic targets in late damages of ionizing radiation, gene expression profile was analyzed in the peripheral blood of three 60Co γ-ray accidently-exposed persons. The results showed that there were 285 up-regulated and 446 down-regulated genes in irradiated samples compared with control samples. The large majority of those differentially expressed genes encode proteins that are associated with immune response, inflammation, cell structure, oxidative stress, neuro-hormone regulation, reproduction, susceptibility of psychiatric disorders, and transcriptional regulators. The expressions of IL3, KDR, CEACAM8 and OSM were validated by real-time RT-PCR method. The findings of our study should help us understanding the molecular mechanisms underlying the late effects of ionizing radiation and to develop better diagonostic and therapeutic strategies for those damages. Fresh blood samples were collected from three 60Coγ-Ray accidentally-exposed persons(3 years post irradiation) and three non-irradiated healthy donors. Leucocytes cells were isolated and total RNA was extracted. Four RNA samples, from three accidently-exposed persons and a RNA mixture extracted from a combination of three healthy donors` leucocytes (ratio 1:1:1, each 3.3×106 cells) in EL buffer, were subjected to Agilent gene expression microarray assay.
Project description:In order to identified as candidate biomarkers for early diagonosis or as therapeutic targets in late damages of ionizing radiation, gene expression profile was analyzed in the peripheral blood of three 60Co γ-ray accidently-exposed persons. The results showed that there were 285 up-regulated and 446 down-regulated genes in irradiated samples compared with control samples. The large majority of those differentially expressed genes encode proteins that are associated with immune response, inflammation, cell structure, oxidative stress, neuro-hormone regulation, reproduction, susceptibility of psychiatric disorders, and transcriptional regulators. The expressions of IL3, KDR, CEACAM8 and OSM were validated by real-time RT-PCR method. The findings of our study should help us understanding the molecular mechanisms underlying the late effects of ionizing radiation and to develop better diagonostic and therapeutic strategies for those damages.
Project description:This study aims to compare mRNA expression between irradiated and non-irradiated people’s blood by single-cell RNA-Seq. In this study, the blood was isolated from patients or healthy persons. The Control group of blood were collected from ten different persons without radiation and mixed for RNA-seq. The blood of irradiated patient was collected from a patient who was exposed to radiation.
Project description:As part of our study in understanding the role of SP140 in inflammatory pathways in macrophages, we inhibited SP140 mRNA using siRNA. Peripheral blood mononuclear cells (PBMCs) were obtained from whole blood of healthy donors (from Sanquin Institute Amsterdam or from GSK Stevenage Blood Donation Unit) by Ficoll density gradient (Invitrogen). CD14+ monocytes were positively selected from PBMCs using CD14 Microbeads according to the manufacturer’s instructions (Miltenyi Biotec). CD14+ cells were differentiated with 20 ng/mL of macrophage colony-stimulating factor (M-CSF) (R&D systems) for 3 days followed by 3 days of polarization into classically activated (inflammatory) M1 macrophages (100 ng/mL IFN-γ; R&D systems). M1 macrophages were transfected with siGENOME human smartpool SP140 siRNA or non-targeting scrambled siRNA for 48h with DharmaFECT™ transfection reagents according to manufacturer’s protocol (Dharmacon). The cells were left unstimulated or stimulated with 100 ng/mL LPS (E. coli 0111:B4; Sigma) for 4h (for qPCR) or 24h (for Elisa). The cells were lysed (ISOLATE II RNA Lysis Buffer RLY-Bioline) for RNA extraction.150 ng total RNA was labelled using the cRNA labelling kit for Illumina BeadArrays (Ambion) and hybridized with Ref8v3 BeadArrays (Illumina). Arrays were scanned on a BeadArray 500GX scanner and data were normalized using quantile normalization with background subtraction (GenomeStudio software; Illumina). This submission only contains processed data
Project description:To examine whether the local carbon ion radiotherapy affects the characteristics of the metastatic tumors, the expression profiles of the primary tumors and the lung metastases were studied in a mouse squamous cell carcinoma model by applying local radiotherapy with no irradiation (negative control), gamma-ray irradiation (reference beam), and carbon-ion irradiation. Keywords: mouse, squamous cell carcinoma, primary tumor, lung metastases, radiotherapy, carbon ion, gamma ray A highly metastatic mouse squamous cell carcinoma NR-S1 was implanted into the hind leg of synergetic C3H/HeNrs mice and irradiated with 5 Gy of carbon ion beam. 8 Gy of gamma ray was used as a reference beam. At 2 weeks after the irradiation, the lung tissue was sampled. In order to collect samples of primary tumors, the tumors were implanted in other mice and irradiated in the same manner, and the primary tumors were collected at 1 week after the irradiation. The tumor cells of the primary and metastatic tumors were collected by laser microdissection, and oligonucleotide microarray analysis of the irradiated primary tumors and the metastatic tumors were all performed in comparison to the non-irradiated primary tumor by two-color methods.