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Radiation effects on a 3D cell culture of primary human keratinocytes and the protective effect of N-acetyl-cysteine and the Traditional Chinese Medicine (TCM) Qingre-Liyan decoction

ABSTRACT: We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3-dimensional (3-D) cell cultures of primary human oral keratinocytes. The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy; Irradiated Tissue with 12 Gy pretreated with N-acetylcysteine (ACS); Irradiated Tissue with 12 Gy pretreated with the Traditional Chinese Medicine (TCM) Qingre-Liyan decoction; and Irradiated Tissue with 12 Gy pretreated with 50:50 w/w ACS:TCM (AT) Group 1: Human Prirmary 3-D Cultures of Human Keratinocytes were irradiated with 12 Gy; Group 2: Human Prirmary 3-D Cultures of Human Keratinocytes were pretereated with N-acetylcysteine (ACS) and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with the Traditional Chinese Medicine (TCM), Qingre-Liyan Decostion and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with a 50:50 w/w mixture of ACS:TCM (AT) and then irradiated with 12 Gy. Each sample was done in triplicate.

ORGANISM(S): Homo sapiens

SUBMITTER: Maria Lambros

PROVIDER: E-GEOD-62397 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Radiation effects on a 3D cell culture of primary human keratinocytes and the protective effect of N-acetyl-cysteine (NAC) and the Traditional Chinese Medicine Qingre-Liyan decoction (QYD)

Project description:We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3-dimensional (3-D) cell cultures of primary human oral keratinocytes. The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy; Irradiated Tissue with 12 Gy pretreated with N-acetylcysteine (NAC); Irradiated Tissue with 12 Gy pretreated with the Traditional Chinese Medicine Qingre-Liyan decoction (QYD); and Irradiated Tissue with 12 Gy pretreated with 50:50 w/w NAC-QYD.

2014-10-16 | GSE62397 | GEO

The role of nitric oxide synthase activity in connection with induced resistance against low-dose hyper-radiosensitivity

Project description:The cells harvested for microarray analysis were high-dose rate (HDR) irradiated, low-dose rate (LDR) irradiated or unirradiated T-47D cells. The HDR-irradiated cells were harvested 24h after a dose of 0.3 Gy at 35 Gy/h, a time where hyper-radiosensitivity (HRS) had returned. The HRS-deficient LDR-irradiated cells were harvested 2 months after a dose of 0.3 Gy at 0.3 Gy/h. LDR-irradiated vs. Control (unirradiated) contains 4 biological replicates. HDR- vs. LDR-irradiated contains 3 biological replicates. HDR-irradiated vs. Control contains 4 biological replicates.

2013-04-15 | E-GEOD-41483 | biostudies-arrayexpress

Genome-wide microarray analysis of immortalized human fibroblasts in response to alpha-particle radiation and the radiation induced bystander effect

Project description:The bystander effect from ionizing radiation consists of cellular responses generated from unirradiated cells to the irradiation of their neighbors. The bystander effect can lead to DNA damage and genomic instability in the affected cells. This non-targeted effect of radiation has received attention due to its potential implications for cancer therapy and radiation protection. Although studied extensively, a complete understanding of its molecular mechanism is the subject of ongoing research. While many studies have targeted specific factors which are suggested to be involved in the bystander effect, few have looked at whole genome gene expression in bystander cells. Furthermore, even fewer studies have looked at the expression in noncancerous human cell lines. In this study we have used a genome-wide microarray approach to investigate transcriptional responses in irradiated and bystander immortalized human fibroblasts following 0.1 Gy ?-particle irradiation. Total RNA was isolated from F11hTERT fibroblasts irradiated with 0.1 Gy ?-particles and bystander fibroblasts receiving medium from control (sham irradiated) and irradiated cells (0.1 Gy). RNA was isolated 4, 8 and 26 h after irradiation.

2011-09-13 | E-GEOD-32091 | biostudies-arrayexpress

Acute and long-term responses to radiation in zebrafish

Project description:A set of changes is identified in the transcription profile associated with the long-term, but not the acute, response to radiation exposure. The study was performed in vivo using zebrafish. To study the long-term response, 24 hour post-fertilization embryos were exposed to 0.1 Gy (low dose) or 1.0 Gy (moderate dose) of whole-body gamma radiation and allowed to develop for 16 weeks. Liver mRNA profiles were then analyzed using the Affymetrix microarray platform, with validation by quantitative PCR. To be able to compare this to the acute response, 16-week old adults were exposed at the same doses and analyzed after 4 hours. We used 5 treatment groups: A=non-irradiated control, allowed to develop for 16 weeks; B=low-dose (0.1 Gy) irradiated, allowed to develop for 16 weeks; C=high-dose (1.0 Gy) irradiate, allowed to develop for 16 weeks; D=16 week old adults irradiated at low dose (0.1 Gy); E=16 week old adults irradiate at high dose (1.0 Gy)

2014-04-01 | E-GEOD-46026 | biostudies-arrayexpress

Proteomics on sera of mice following local high-dose heart irradiation

Project description:Background and Purpose: Cardiotoxicity is a well-known adverse effect of radiation therapy. Measurable abnormalities in the heart function indicate advanced and often irreversible heart damage. Therefore, early detection of cardiac toxicity is necessary to delay and alleviate the development of the disease. The present study investigated long-term serum proteome alterations following local heart irradiation using a mouse model with the aim to detect biomarkers of radiation-induced cardiac toxicity. Materials and Methods: Serum samples from C57BL/6J mice were collected 20 weeks after local heart irradiation with 8 Gy or 16 Gy X-ray; the controls were sham-irradiated. The samples were analyzed by quantitative proteomics based on data-independent acquisition mass spectrometry. The proteomics data were further investigated using bioinformatics and ELISA. Results: The analysis showed radiation-induced changes in the level of several serum proteins involved in the acute phase response, inflammation and cholesterol metabolism. We found significantly enhanced expression of pro-inflammatory cytokines (TNF-, TGF-, IL-1 and IL-6) in the serum of the irradiated mice. The level of free fatty acids, total cholesterol, low density lipoprotein (LDL) and oxidized LDL was increased whereas that of high density lipoprotein was decreased by irradiation. Conclusions: This study provides information on systemic effects of heart irradiation. It elucidates a radiation fingerprint in the serum that may be used to elucidate adverse cardiac effects after radiation therapy.

2021-09-10 | PXD024446 | Pride

p38a-dependent gene expression in keratinocytes

Project description:Gene expression in wild-type and p38a-knockout keratinocytes were compared. Keratinocytes were isolated from newborn mice, and left unirradiated (0 h) and irradiated (4 h) with ultraviolet-B (UVB). C57BL/6 wild-type mice, and keratinocyte-specific p38a-knockout mice on a C57BL/6 background were used for isolation of primary keratinocytes. Gene expression in keratinocytes was analyzed 0 and 4 h after UVB irradiation (75 mJ/cm2).

2014-01-01 | E-GEOD-51206 | biostudies-arrayexpress

Expression data from human unbilical endothelial cells (HUVEC) irradiated with X-ray

Project description:To understand the molecular mechanism underlying inflammatory reaction in vascular system post exposure to ionizing radiation, we carried out microarray analysis in HUVEC exposed with X-ray HUVEC were irradiated with X-ray (2.5 Gy) and then cultured for 6, 12, and 24 hr. Total RNA were extracted from the tissue by QIAGEN Rneasy mini kit.

2016-01-21 | E-GEOD-76484 | biostudies-arrayexpress

Genome-wide microarray analysis of normal human fibroblasts in response to γ-radiation and the radiation induced bystander effect

Project description:The bystander effect from ionizing radiation consists of cellular responses generated from non-irradiated cells to the irradiation of their neighbors. The bystander effect is predominant at low doses and can lead to DNA damage and genomic instability in the affected cells. This non-targeted effect of radiation has received attention due to its potential implications for cancer therapy and radiation protection. Although studied extensively, a complete understanding of its molecular mechanism is the subject of ongoing research. While many studies have targeted specific factors which are suggested to be involved in the bystander effect, few have looked at whole genome gene expression in bystander cells. Furthermore, even fewer studies have looked at the expression in normal human cell lines. In this study, we have monitored transcriptional responses to γ-radiation in irradiated and bystander normal fibroblasts simultaneously using a genome-wide microarray approach. Bystander fibroblasts incubated in medium from irradiated cells, showed transient enrichment (less than 1.5 fold) in ribosome and oxidative phosphorylation pathways, and neurodegenerative disease pathways associated with mitochondrial dysfunctions. Bystander fibroblasts did not, however, display increases in oxidative stress, a phenomenon often linked with the radiation induced bystander effect. Total RNA was isolated from normal human fibroblasts irradiated with 2.0 Gy and fibroblasts incubated with medium from sham irradiated and irradiated cells 2 h after irradiation. RNA was isolated 4, 8 and 26 h after irradiation and there are 4 replicates for each sample for a total of 36 samples.

2010-12-03 | E-GEOD-25772 | biostudies-arrayexpress

Expression data from 5 human cell lines exposed to IR (5 Gy)

Project description:The cellular response to DNA damage is vital for maintaining genomic stability and preventing undue cell death or cancer formation. The DNA damage response (DDR), most robustly mobilized by double-strand breaks (DSBs), rapidly activates an extensive signaling network that affects numerous cellular systems, leading to cell survival or programmed cell death. A major component of the DDR is the widespread modulation of gene expression. We analyzed transcriptional responses to ionizing radiation (IR) in 5 human cell lines to elucidate the scope of this response and identify its gene targets. According to the mRNA expression profiles most of the responses were cell line-specific. Data analysis identified significant enrichment for p53 target genes and cell cycle-related pathways among groups of up-regulated and down-regulated genes, respectively. Expression profiles were measured using affymetrix chips in IR- irradiated G361 cells and their time-matched untreated controls. Time points recorded were 0, 3 and 6 hrs. IR dose: 5 Gy Expression profiles were measured using affymetrix chips in IR- irradiated HepG2 cells and their time-matched untreated controls. Time points recorded were 0, 3 and 6 hrs. IR dose: 5 Gy Expression profiles were measured using affymetrix chips in IR- irradiated TK6 cells and their time-matched untreated controls. Time points recorded were 0, 3 and 6 hrs. IR dose: 5 Gy Expression profiles were measured using affymetrix chips in IR- irradiated U2OS cells and their time-matched untreated controls. Time points recorded were 0, 3 and 6 hrs. IR dose: 5 Gy Expression profiles were measured using affymetrix chips in IR- irradiated BJ cells and their time-matched untreated controls. Time points recorded were 0, 3 and 6 hrs. IR dose: 5 Gy

2011-07-31 | E-GEOD-30240 | biostudies-arrayexpress

Response of human lymphoblastoid cells to HZE (iron ions) or gamma-rays

Project description:Transcriptional profiling of human lymphoblastoid TK6 cells comparing mock irradiated cells with cells exposed 24 hours previously to 1.67 Gy HZE (1 GeV/amu iron ions accelerated at the NASA Space Research Laboratory (NSRL) of Brookhaven National Laboratory) or 2.5 Gy 137Cs gamma rays. TK6 cells were mock irradiated or exposed to HZE or gamma-rays, and RNA was harvested 24 hours later. 3 biological replicates were independently grown and harvested during three different runs at the NSRL. One replicate per array.

2011-04-01 | E-GEOD-16518 | biostudies-arrayexpress

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