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Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray.

Publication ,  Journal Article
Braam, B; Allen, P; Benes, E; Koomans, HA; Navar, LG; Hammond, T
Published in: Eur J Pharmacol
March 19, 2003

Angiotensin II has been shown to exert complex effects on proximal tubular cell function and growth. To assess some of the direct effects on proximal tubular cells, changes in gene expression of selected cellular pathways were determined after exposure to angiotensin II. We used DNA microarrays to analyze multiple gene expression responses to increasing angiotensin II concentrations. Human proximal tubular cells were grown in flasks, and the presence of angiotensin type 1 receptor was confirmed by Western blot analysis. At passages 4-6, these cells were exposed to angiotensin II and harvested 4 h later and mRNA of the cells was extracted; 2 microg of mRNA was fluorescently conjugated for cDNA microarray hybridization. A custom-made DNA microarray was designed by selecting 300 human genes from 10 different functional systems and amplifying clones using polymerase chain reaction. Cells were subjected to 10 and 100 nM angiotensin II with paired untreated cells as controls. RNA was isolated, reverse transcribed, labeled and hybridized to the arrays and the ratios calculated. Ratios of > or =2.0 and < or =0.5 were considered significant. Coordinated changes were observed in genes of the hepatocyte nuclear factor 3 family (NHF3; HNF3A, HNF3B and HNF3G), in the E2F genes (E2F1, E2F3) and the interferon regulatory factors IRF1 and IRF5. Induction of the expression of transcription factors points towards complex regulation of gene expression upon angiotensin II exposure. Three genes involved in the dampening of oxidative stress were enhanced. Taken together, brief exposure of human tubular epithelial cells to angiotensin II elicited a marked induction of nuclear factors, antioxidant genes and hormones and hormone receptor genes. The quick activation of transcription factors by angiotensin II indicates that angiotensin II can directly initiate a cascade of expressional events in proximal tubular cells.

Duke Scholars

Published In

Eur J Pharmacol

DOI

ISSN

0014-2999

Publication Date

March 19, 2003

Volume

464

Issue

2-3

Start / End Page

87 / 94

Location

Netherlands

Related Subject Headings

  • Reverse Transcriptase Polymerase Chain Reaction
  • Receptors, Angiotensin
  • Receptor, Angiotensin, Type 1
  • RNA, Messenger
  • Pharmacology & Pharmacy
  • Oligonucleotide Array Sequence Analysis
  • Kidney Tubules, Proximal
  • Humans
  • Gene Expression Regulation
  • Dose-Response Relationship, Drug
 

Citation

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Braam, B., Allen, P., Benes, E., Koomans, H. A., Navar, L. G., & Hammond, T. (2003). Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray. Eur J Pharmacol, 464(2–3), 87–94. https://doi.org/10.1016/s0014-2999(03)01382-7
Braam, Branko, Pat Allen, Ed Benes, Hein A. Koomans, L Gabriel Navar, and Timothy Hammond. “Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray.Eur J Pharmacol 464, no. 2–3 (March 19, 2003): 87–94. https://doi.org/10.1016/s0014-2999(03)01382-7.
Braam B, Allen P, Benes E, Koomans HA, Navar LG, Hammond T. Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray. Eur J Pharmacol. 2003 Mar 19;464(2–3):87–94.
Braam, Branko, et al. “Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray.Eur J Pharmacol, vol. 464, no. 2–3, Mar. 2003, pp. 87–94. Pubmed, doi:10.1016/s0014-2999(03)01382-7.
Braam B, Allen P, Benes E, Koomans HA, Navar LG, Hammond T. Human proximal tubular cell responses to angiotensin II analyzed using DNA microarray. Eur J Pharmacol. 2003 Mar 19;464(2–3):87–94.
Journal cover image

Published In

Eur J Pharmacol

DOI

ISSN

0014-2999

Publication Date

March 19, 2003

Volume

464

Issue

2-3

Start / End Page

87 / 94

Location

Netherlands

Related Subject Headings

  • Reverse Transcriptase Polymerase Chain Reaction
  • Receptors, Angiotensin
  • Receptor, Angiotensin, Type 1
  • RNA, Messenger
  • Pharmacology & Pharmacy
  • Oligonucleotide Array Sequence Analysis
  • Kidney Tubules, Proximal
  • Humans
  • Gene Expression Regulation
  • Dose-Response Relationship, Drug