J Korean Soc Ther Radiol Oncol.  2006 Mar;24(1):58-66.

Radiation Induced Changes in the Expression of Fibronectin, Pai-1, MMP in Rat Glomerular Epithelial Cell

• Park WY
• Kim WD
• Zheng Y
• Ha TS
• Kim JS
• Cho MJ

Abstract

PURPOSE: Renal irradiation can lead to the development of radiation nephropathy, and this is characterized by the accumulation of extracellular matrix and final fibrosis. To determine the possible role of the glomerular epithelial cell, the radiation-induced changes in the expression of its genes associated with the extracellular matrix were analyzed.
MATERIALS AND METHODS
Rat glomerular epithelial cells (GEpC) were irradiated with a single dose of 0, 2, 5, 10 and 20 Gy with using 6 MV LINAC (Siemens, USA), and the samples were collected 6, 24, 48 and 72 hours post-irradiation, respectively. Northern blotting, western blotting and zymography were used to measure the expression level of fibronectin (Fn), plasminogen activator inhibitor-1 (Pai-1), matrix metalloproteinases-2, 9 (MMP-2, 9), tissue inhibitor of metalloproteinase-2 (TIMP-2), tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA).
RESULTS
Irradiation with a single dose of 10 Gy resulted in a significant increase in Fn mRNA since 24 hours post-irradiation, and a single dose of 5 and 10 Gy significantly increased the Fn immunoreactive protein measured 48 hours post-irradiation. An increase in Pai-1 mRNA and protein was also observed and especially, a single dose of 10 Gy significantly increased the mRNA measured 24 and 48 hours post-irradiation. The active MMP-2 measured 24 hours post-irradiation slightly increased in a dose dependent manner, but this increase did not reach statistical significance. The levels of MMP-9, TIMP-2, t-PA and u-PA appeared unaltered after irradiation.
CONCLUSION
Irradiation of the glomerular epithelial cells altered the expression of genes associated with the extracellular matrix, implying that the glomerular epithelial cell may be involved in the development of radiation nephropathy.

Keyword

Rat glomerular epithelial cell; Radiation; Fibronectin; Pai-1; MMP

MeSH Terms

Animals
Blotting, Northern
Blotting, Western
Epithelial Cells*
Extracellular Matrix
Fibronectins*
Fibrosis
Plasminogen Activator Inhibitor 1*
Plasminogen Activators
Rats*
RNA, Messenger
Tissue Inhibitor of Metalloproteinase-2
Tissue Plasminogen Activator
Urokinase-Type Plasminogen Activator
Fibronectins
Plasminogen Activator Inhibitor 1
Plasminogen Activators
RNA, Messenger
Tissue Inhibitor of Metalloproteinase-2
Tissue Plasminogen Activator
Urokinase-Type Plasminogen Activator