Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human mono or co-cultured HMEC-1 and HK-2 cells to identify the influence of microvascular endothelial cells on transcriptional regulation of proximal tubular epithelial cells

ABSTRACT: The effect of the presence of HMEC-1 cells on HK-2 cell gene expression was investigated. Cells were cultured in mono or co-culture on opposite sides of aluminium oxide filters, 0.2 ?µM pore size. These conditions provide a close proximity but not direct contact. RNA from HK-2 cells was assayed with Affymetrix HGU133 Plus 2 arrays. 3 Biological samples were used per group.

ORGANISM(S): Homo sapiens

SUBMITTER: Paul Jennings

PROVIDER: E-MEXP-1178 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Influence of microvascular endothelial cells on transcriptional regulation of proximal tubular epithelial cells.

Aydin Sonia S Signorelli Sara S Lechleitner Thomas T Joannidis Michael M Pleban Clara C Perco Paul P Pfaller Walter W Jennings Paul P

American journal of physiology. Cell physiology 20071205 2

In the renal cortex the peritubular capillary network and the proximal tubular epithelium cooperate in solute and water reabsorption, secretion, and inflammation. However, the mechanisms by which these two cell types coordinate such diverse functions remain to be characterized. Here we investigated the influence of microvascular endothelial cells on proximal tubule cells, using a filter-based, noncontact, close-proximity coculture of the human microvascular endothelial cell line HMEC-1 and the h ...[more]

PMID: 18057119

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Project description:Aim： Use microarray analysis to understand the molecular mechanism underlying the effect of aristolochic acid (AA), a major active component of plants from the Aristolochiaceae family, in normal human kidney (HK-2) cells. Methods： HK-2 cells were treated with AA for 24 hours and cell viability was measured by a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay. Complementary DNA microarrays were used to investigate the gene expression pattern of HK-2 cells exposed to AA and the results of this study were in triplicate. Quantitative real-time RT-PCR assay was used to verify the microarray data for selected nuclear factor kappa B (NF-κB)-regulated genes. Furthermore, subcellular localization of NF-κB p65 was visualized by immunofluorescence confocal microscopy in HK-2 cells. NF-κB activity was examined by luciferase reporter assay in HK-2/NF-κB transgenic cells. Results： AA exhibited a dose-dependent cytotoxic effect in HK-2 cells and induced alterations in gene expression profiles related to DNA damage response, stress response, etc. In addition, 9 biological pathways associated with immunomodulatory functions were down-regulated in AA-treated HK-2 cells. Network analysis revealed that NF-κB played a central role in the network topology. Among NF-kB-regulated genes, 8 differentially expressed genes were verified by real-time RT-PCR. The inhibition of NF-κB activity by AA was further confirmed by immunofluorescence confocal microscopy and by NF-κB luciferase reporter assay. Conclusion： Our data revealed that AA could suppress NF-κB activity in normal human cells, perhaps partially accounting for the reported anti-inflammatory effects of some plants from the genus Aristolochia. HK-2 cells were grown in keratinocyte serum-free basal medium (Gibco) supplemented with 5 ng/ml of recombinant epidermal growth factor and 50 μg/ml of bovine pituitary extract without antibiotics in 5 % CO2 at 370C. HK-2 cells were seeded in 25-T flasks and incubated for 24 h before aristolochic acid treatment. Aristolochic acid (10 90 μM) were added to HK-2 cells for 24 h. The control cells received equal amounts of water only.

Dataset Information

Transcription profiling of human mono or co-cultured HMEC-1 and HK-2 cells to identify the influence of microvascular endothelial cells on transcriptional regulation of proximal tubular epithelial cells

Publications

Influence of microvascular endothelial cells on transcriptional regulation of proximal tubular epithelial cells.

Similar Datasets

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