Limits...
Transcriptome analysis of human OXR1 depleted cells reveals its role in regulating the p53 signaling pathway.

Yang M, Lin X, Rowe A, Rognes T, Eide L, Bjørås M - Sci Rep (2015)

Bottom Line: The oxidation resistance gene 1 (OXR1) is crucial for protecting against oxidative stress; however, its molecular function is unknown.In total, in non-treated and hydrogen peroxide exposed HeLa cells, OXR1 depletion resulted in down-regulation of 554 genes and up-regulation of 253 genes.In summary, OXR1 may act as a sensor of cellular oxidative stress to regulate the transcriptional networks required to detoxify reactive oxygen species and modulate cell cycle and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Oslo University Hospital and University of Oslo, Norway.

ABSTRACT
The oxidation resistance gene 1 (OXR1) is crucial for protecting against oxidative stress; however, its molecular function is unknown. We employed RNA sequencing to examine the role of human OXR1 for genome wide transcription regulation. In total, in non-treated and hydrogen peroxide exposed HeLa cells, OXR1 depletion resulted in down-regulation of 554 genes and up-regulation of 253 genes. These differentially expressed genes include transcription factors (i.e. HIF1A, SP6, E2F8 and TCF3), antioxidant genes (PRDX4, PTGS1 and CYGB) and numerous genes of the p53 signaling pathway involved in cell-cycle arrest (i.e. cyclin D, CDK6 and RPRM) and apoptosis (i.e. CytC and CASP9). We demonstrated that OXR1 depleted cells undergo cell cycle arrest in G2/M phase during oxidative stress and increase protein expression of the apoptosis initiator protease CASP9. In summary, OXR1 may act as a sensor of cellular oxidative stress to regulate the transcriptional networks required to detoxify reactive oxygen species and modulate cell cycle and apoptosis.

No MeSH data available.


Related in: MedlinePlus

Validation of RNA-seq data by qPCR.(a,b) Genes involved in p53 signaling pathway. (c,d) Transcription factors. (e,f) Early stress response genes. (a,c,e) Data extracted from RNA-seq. (b,d,f) The mRNA levels of the same set of genes were measured by qPCR. The mRNA level was presented as the fold change as compared to non-treated control cells. The standard deviation was calculated from 4 cDNA samples measured in duplicate. siCon: control siRNA; siOXR1: hOXR1 siRNA;. NT: non-treatment; R0h: cells treated with H2O2 0.5 mM for 1 h and harvested immediately without recovery. *p < 0.01 compared to siCon_NT; #p < 0.01 compared to siCon_R0h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4663793&req=5

f4: Validation of RNA-seq data by qPCR.(a,b) Genes involved in p53 signaling pathway. (c,d) Transcription factors. (e,f) Early stress response genes. (a,c,e) Data extracted from RNA-seq. (b,d,f) The mRNA levels of the same set of genes were measured by qPCR. The mRNA level was presented as the fold change as compared to non-treated control cells. The standard deviation was calculated from 4 cDNA samples measured in duplicate. siCon: control siRNA; siOXR1: hOXR1 siRNA;. NT: non-treatment; R0h: cells treated with H2O2 0.5 mM for 1 h and harvested immediately without recovery. *p < 0.01 compared to siCon_NT; #p < 0.01 compared to siCon_R0h.

Mentions: In order to verify the role of hOXR1 in gene expression regulation of the p53 signaling pathway, we measured mRNA levels of the differentially expressed genes in the p53 pathway using Real-Time quantitative PCR (qPCR). Consistent with the RNA-seq data, the qPCR data showed significant differential expression of all genes tested (Fig. 4a,b). For hOXR1, its expression in mRNA level was significantly reduced in hOXR1 siRNA transfected cells (less than 10% expression as compared to control cells (siCon) (Fig. 4a,b).


Transcriptome analysis of human OXR1 depleted cells reveals its role in regulating the p53 signaling pathway.

Yang M, Lin X, Rowe A, Rognes T, Eide L, Bjørås M - Sci Rep (2015)

Validation of RNA-seq data by qPCR.(a,b) Genes involved in p53 signaling pathway. (c,d) Transcription factors. (e,f) Early stress response genes. (a,c,e) Data extracted from RNA-seq. (b,d,f) The mRNA levels of the same set of genes were measured by qPCR. The mRNA level was presented as the fold change as compared to non-treated control cells. The standard deviation was calculated from 4 cDNA samples measured in duplicate. siCon: control siRNA; siOXR1: hOXR1 siRNA;. NT: non-treatment; R0h: cells treated with H2O2 0.5 mM for 1 h and harvested immediately without recovery. *p < 0.01 compared to siCon_NT; #p < 0.01 compared to siCon_R0h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663793&req=5

f4: Validation of RNA-seq data by qPCR.(a,b) Genes involved in p53 signaling pathway. (c,d) Transcription factors. (e,f) Early stress response genes. (a,c,e) Data extracted from RNA-seq. (b,d,f) The mRNA levels of the same set of genes were measured by qPCR. The mRNA level was presented as the fold change as compared to non-treated control cells. The standard deviation was calculated from 4 cDNA samples measured in duplicate. siCon: control siRNA; siOXR1: hOXR1 siRNA;. NT: non-treatment; R0h: cells treated with H2O2 0.5 mM for 1 h and harvested immediately without recovery. *p < 0.01 compared to siCon_NT; #p < 0.01 compared to siCon_R0h.
Mentions: In order to verify the role of hOXR1 in gene expression regulation of the p53 signaling pathway, we measured mRNA levels of the differentially expressed genes in the p53 pathway using Real-Time quantitative PCR (qPCR). Consistent with the RNA-seq data, the qPCR data showed significant differential expression of all genes tested (Fig. 4a,b). For hOXR1, its expression in mRNA level was significantly reduced in hOXR1 siRNA transfected cells (less than 10% expression as compared to control cells (siCon) (Fig. 4a,b).

Bottom Line: The oxidation resistance gene 1 (OXR1) is crucial for protecting against oxidative stress; however, its molecular function is unknown.In total, in non-treated and hydrogen peroxide exposed HeLa cells, OXR1 depletion resulted in down-regulation of 554 genes and up-regulation of 253 genes.In summary, OXR1 may act as a sensor of cellular oxidative stress to regulate the transcriptional networks required to detoxify reactive oxygen species and modulate cell cycle and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Oslo University Hospital and University of Oslo, Norway.

ABSTRACT
The oxidation resistance gene 1 (OXR1) is crucial for protecting against oxidative stress; however, its molecular function is unknown. We employed RNA sequencing to examine the role of human OXR1 for genome wide transcription regulation. In total, in non-treated and hydrogen peroxide exposed HeLa cells, OXR1 depletion resulted in down-regulation of 554 genes and up-regulation of 253 genes. These differentially expressed genes include transcription factors (i.e. HIF1A, SP6, E2F8 and TCF3), antioxidant genes (PRDX4, PTGS1 and CYGB) and numerous genes of the p53 signaling pathway involved in cell-cycle arrest (i.e. cyclin D, CDK6 and RPRM) and apoptosis (i.e. CytC and CASP9). We demonstrated that OXR1 depleted cells undergo cell cycle arrest in G2/M phase during oxidative stress and increase protein expression of the apoptosis initiator protease CASP9. In summary, OXR1 may act as a sensor of cellular oxidative stress to regulate the transcriptional networks required to detoxify reactive oxygen species and modulate cell cycle and apoptosis.

No MeSH data available.


Related in: MedlinePlus