Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of RNA. The goals of this study are to compare HuR binding RNAs between 0 or 5 Gy irradiation, and analyze its effect on RNA expression. Methods: Human skin fibroblast cell line WS1 cells were treated with 0 or 5 Gy irradiation. After 24 h, RNA expression was measured using using Illumina HiSeq4000. Meanwhile, RIP was performed using HuR antibody. HuR bound mRNAs after 0 or 5 Gy irradiation was measured using using Illumina HiSeq4000. Results: RIP-sequencing reveals HuR binding RNAs between 0 or 5 Gy irradiation in WS1 cells. RNA sequencing reveals dyeregulated RNAs between 0 and 5 Gy irradiation. Conclusions: Our study represents the first detailed analysis of HuR binding RNAs between 0 and 5 Gy irradiation generated by RNA-seq technology. This study contributes to our understanding of the role of HuR inresponse to radiation.
Project description:Breast cancer ranks top in the incidence among the main sites of female cancer in Japan. The epidemiological study on atomic bomb survivors has suggested that the excess relative risk for breast cancer is higher than any other sites. Little is known, however, about the molecular mechanisms of breast cancer induction by radiation. Therefore, we analyzed here the genome-wide copy number aberration of radiation-induced rat mammary carcinomas using microarray-based comparative genomic hybridization (array-CGH). Mammary carcinomas were induced by 2 Gy gamma irradiation of Sprague-Dawley (SD) rats at 3 or 7 weeks of age. We examined 14 mammary carcinomas induced by gamma-irradiation (2 Gy) and found 26 aberrations including trisomies of chromosomes 4 and 10 in 3 and 1 carcinomas, respectively, and deletion of chromosomes 3q35q36 and 5q32 (Cdkn2a and Cdkn2b region) in 2 and 2 carcinomas, respectively. On the other hand, only one aberration (amplification of chromosome 10q31) was observed in four spontaneous mammary carcinomas. These results suggest that the trisomy of chromosome 4 and deletion of chromosomes 3q35q36 and 5q32 were associated with radiation exposure.
Project description:Here, we present a whole-genome survey of the murine transcriptomic response to physiologically-relevant radiation doses, 2 and 8 Gy. There are 18 distinct biological samples here. Mice were exposed to ionizing radiation (Cesium-138 source) and whole blood was collected by cardiac puncture 6 hours post treatment. Doses were 0 (7 samples), 2 (5 samples), and 8 (6 samples) gy.
Project description:small RNA seq for miRNA differential expression analysis were performed in primary human aotic endothelial cells treated with increasing doses of radiation (1 Gy, 2 Gy, 4 Gy, 8 Gy and 10 Gy). Total RNA was collected 24 h and 72 h after radiation. Comparisons were made with unirradiated cells plated for 24 h and 72 h repsectively.
Project description:The Russian part of C-HPP is aimed to identify proteins encoded by the 18 human chromosome. In this project, 3 liver tissue samples were studied by standart Shotgun and 2 dimensional liquid chromatography approach to enhance the sensitivity of the MS method. The total amount of proteins found in samples is 5022 and the proteins encoded by the 18 human chromosome is 64.
Project description:The Russian part of C-HPP is aimed to identify proteins encoded by the 18 human chromosome. In this project, 3 liver tissue samples were studied by standart SRM and 2 dimensional liquid chromatography approach to enhance the sensitivity of the MS method. The total amount of proteins found in samples encoded by the 18 human chromosome is 118. UPS1 used as calibration and to measure the sensitivity of SRM method.
Project description:HaCaT cell line is a kind of keratinocyte from human skin. We used single cell RNA sequencing (scRNA-seq) to analyze the influnce of fractionated 2 Gy or single 20 Gy irradiation on HaCaT cells.
Project description:To investigate DNA copy number changes in chromosome 18 of small intestinal tumors in B6/B6-Chr18MSM-F1 ApcMin/+ mice irradiated with ionizing radiation
Project description:Background: Patients with small intestinal neuroendocrine tumors (SINETs) frequently present with lymph node and liver metastases at the time of diagnosis, but the molecular changes that lead to the progression of these tumors are largely unknown. Sequencing studies have only identified recurrent point mutations in a single gene, CDKN1B, with heterozygous mutations in less than 10% of all tumors. Although SINETs are genetically stable tumors with a low frequency of point mutations and indels, they often harbor recurrent hemizygous copy number alterations (CNAs) yet the functional implications of these CNA are unclear. Methods: Utilizing comparative genomic hybridization (CGH) arrays we analyzed the CNA profile of 131 SINETs from 117 patients. Two tumor suppressor genes and corresponding proteins i.e. SMAD4, and CDKN1B, were further characterized using a tissue microarray (TMA) with 846 SINETs. Immunohistochemistry (IHC) was used to quantify protein expression in TMA samples and this was correlated with chromosome number evaluated with fluorescent in-situ hybridization (FISH). Intestinal tissue from a Smad4+/- mouse model was used to detect entero-endocrine cell hyperplasia with IHC. Results: Analyzing the CGH arrays we found loss of chromosome 18q and SMAD4 in 71% of SINETs and that focal loss of chromosome 12 affecting the CDKN1B was present in 9.4% of SINETs. No homozygous loss of chromosome 18 was detected. Hemizygous loss of SMAD4, but not CDKN1B, significantly correlated with reduced protein levels but hemizygous loss of SMAD4 did not induce entero-endocrine cell hyperplasia in the Smad4+/- mouse model. Conclusions: Hemizygous loss of chromosome 18q and the SMAD4 gene is the most common genetic event in SINETs and our results suggests that this could influence SMAD4 protein expression. Although SMAD4 haploinsufficiency alone did not induce tumor initiation, loss of chromosome 18 could represent an evolutionary advantage in SINETs explaining the high prevalence of this aberration. Functional consequences of reduced SMAD4 protein levels could hypothetically be a potential mechanism as to why loss of chromosome 18 appears to be clonally selected in SINETs.
Project description:Utilizing an established model of Radiation Induced Pulmonary Fibrosis, low input RNA sequencing was performed on whole lung cell suspensions obtained from 12.5 Gy thorax only radiation treated C57BL/6J mice and compareed to 0 Gy (Sham irradiation) age matched controls