Expression and Regulation of Latent TGF-beta Binding Protein-1 Transcripts and Their Splice Variants in Human Glomerular Endothelial Cells.
10.3346/jkms.2005.20.4.628
- Author:
Joon Hyeok KWAK
1
;
Ji Su WOO
;
Kunyoo SHIN
;
Hee Joon KIM
;
Hoe Su JEONG
;
Dong Cheol HAN
;
Sung Il KIM
;
Choon Sik PARK
Author Information
1. Hyonam Kidney Laboratory, Soon Chun Hyang University, Korea. hblee@hkl.ac.kr
- Publication Type:Original Article ; Comparative Study ; Research Support, Non-U.S. Gov't
- Keywords:
Latent TGF-beta binding protein;
Splice Variant of LTBP-1;
Endothelial Cells;
Trnasforming growth factor beta1;
Blood Glucose;
Hydrogen Peroxide;
Vascular Endothelial Growth Factor
- MeSH:
*Alternative Splicing;
Amino Acid Sequence;
Cell Line;
Cells, Cultured;
Cloning, Molecular;
Comparative Study;
Endothelial Cells/drug effects/*metabolism;
*Gene Expression Regulation;
Glucose/pharmacology;
Humans;
Hydrogen Peroxide/pharmacology;
Intracellular Signaling Peptides and Proteins/*genetics;
Kidney Glomerulus/cytology;
Protein Isoforms/genetics;
RNA, Messenger/genetics/metabolism;
Research Support, Non-U.S. Gov't;
Reverse Transcriptase Polymerase Chain Reaction;
*Transcription, Genetic;
Transfection;
Transforming Growth Factor beta/pharmacology;
Vascular Endothelial Growth Factor A/pharmacology
- From:Journal of Korean Medical Science
2005;20(4):628-635
- CountryRepublic of Korea
- Language:English
-
Abstract:
Latent transforming growth factor (TGF)-beta-binding protein (LTBP) is required for the assembly, secretion, matrix association, and activation of latent TGF-beta complex. To elucidate the cell specific expression of the genes of LTBP-1 and their splice variants and the factors that regulate the gene expression, we cultured primary human glomerular endothelial cells (HGEC) under different conditions. Basal expression of LTBP-1 mRNA was suppressed in HGEC compared to WI-38 human embryonic lung fibroblasts. High glucose, H2O2, and TGF-beta1 upregulated and vascular endothelial growth factor (VEGF) further downregulated LTBP-1 mRNA in HGEC. RT-PCR with a primer set for LTBP-1S produced many clones but no clone was gained with a primer set for LTBP-1L. Of 12 clones selected randomly, Sca I mapping and DNA sequencing revealed that only one was LTBP-1S and all the others were LTBP-1S delta 53. TGF-beta1, but not high glucose, H2O2 or VEGF, tended to increase LTBP-1S delta 53 mRNA. In conclusion, HGEC express LTBP-1 mRNA which is suppressed at basal state but upregulated by high glucose, H2O2, and TGF-beta1 and downregulated by VEGF. Major splice variant of LTBP-1 in HGEC was LTBP-1S delta 53. Modification of LTBP-1S delta 53 gene in HGEC may abrogate fibrotic action of TGF-beta1 but this requires confirmation.
