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Fibronectin expression in human mesangial cell cultures and its alterations by adriamycin

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Abstract

Human mesangial cell (HMC) cultures synthesize cellular fibronectin (FN), which is secreted and incorporated into a fibrillar extracellular matrix (ECM). The anticancer drug adriamycin (ADR) induces changes in extracellular FN deposition. As revealed by immunofluorescence staining, a 24 h incubation of the cells with 2 μg ADR/ml resulted in a marked expansion of the pericellular FN fibers, which may be due to either an increased synthesis or a decreased FN degradation. The effects of ADR on FN mRNA were analysed by northern hybridization andin vitro translation. Steady-state FN mRNA levels were significantly increased by 60% following ADR administration. However, yields of radioactivity incorporated into FN by cell-free translation remained constant (2.3±0.7%,n=24, vs controls 2.2±0.8% of total radioactivity,n=23). The quality of translation products was not affected by the drug, whereas translation efficiency of total RNA from ADR-treated HMC was only 75% of controls. The data presented suggest a negative feedback control of FN expression on the level of translation. Extracellular FN accumulation in the experimental model of ADR-induced progressive glomerulopathy therefore cannot be explained by an increased FN synthesis, but is rather regarded a consequence of proteinase inhibition. This assumption is compatible with a diminished number of FN fragments recently demonstrated in the culture medium of ADR-treated HMC, and is further corroborated by the loss of urinary FN degradation products accompanying the onset of proteinuria in ADR-treated rats.

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Abbreviations

ADR:

adriamycin

BSA:

boyine serum albumin

ECM:

extracellular matrix

EDTA:

ethylenediamine tetraacetic acid

FITC:

fluorescein isothiocyanate

FN:

fibronectin

HMC:

human mesangial cell

PBS:

phosphate buffered saline

PMSF:

phenylmethylsulfonyl fluoride

SDS:

sodium dodecyl sulfate

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

SSC:

standard saline citrate

SSPE:

standard saline phosphate ethylenediamine tetraacetic acid disodium salt

TGF-β:

transforming growth factor β

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Authors and Affiliations

  1. Institute of Animal Physiology, Justus-Liebig University Giessen, Germany

    M. Soose, A. Padur & D. Oberst

  2. Division of Nephrology, Hannover Medical School, Hannover, Germany

    S. Wenzel & H. Stolte

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  1. M. Soose
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  2. S. Wenzel
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  3. A. Padur
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  4. D. Oberst
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  5. H. Stolte
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Soose, M., Wenzel, S., Padur, A. et al. Fibronectin expression in human mesangial cell cultures and its alterations by adriamycin. Cell Biol Toxicol 11, 51–63 (1995). https://doi.org/10.1007/BF00769992

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  • DOI: https://doi.org/10.1007/BF00769992

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