Abstract: SA-PO567
D-Site Binding Protein Regulates Cell Proliferation Through Mediating Cell Cycle Progression in Rat Mesangial Cells
Session Information
- Glomerular Diseases: Fibrosis, Extracellular Matrix
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1201 Glomerular Diseases: Fibrosis and Extracellular Matrix
Authors
- Chen, Lei, Xi'an Jiaotong University, Xi'an, China
- Li, Jie, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, China
- Qu, Ning, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, China
- Liu, Hua, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, China
Background
Over proliferation of glomerular mesangial cells (MCs) disturbs mesangial homeostasis and leads to renal damage in mesangioproliferative glomerulonephritis. It is documented that transcriptional factors may be involved in the proliferation of MCs. This study aims to identify the key transcriptional factor that prevents the MCs from over proliferation and to clarify its regulatory mechanism.
Methods
Glomeruli were isolated from rats subjected to anti-Thy1 antibody or phosphate-buffered saline. Total RNA was extracted and subjected to microarray analysis. Lentiviral transfection and siRNA transfection were used to obtain D-site binding protein (DBP)-over expressed and DBP-knockdown rat primary MCs, respectively. The cell proliferative capacity was measured by 5-Ethynyl-2′-deoxyuridine (EdU) assay. Flow cytometry was conducted for cell cycle analysis. Histology, immunohistochemistry, and western blot analysis were performed to detect the expression of selected gene and cell cycle regulators.
Results
The cell cycle pathway was the most enriched pathway in the anti-Thy1N model, and the DBP ranked first in the cluster of transcription factors, which was markedly decreased accompanied by an over proliferation of MCs in anti-Thy1N model rats. The knockdown of DBP significantly promoted the proliferation of primary rat MCs, whereas the DBP over expression inhibited the MCs’ proliferative capacity in vitro, compared to that of the control cells. DBP arrested G1/S-phase transition by inhibiting the expression of p21, p27 and inducing the Cyclin D1 expression in MCs.
Conclusion
DBP effectively inhibits the proliferation of MCs through G0/G1 phase arrest, and the decrease of DBP may induce mesangial over proliferation in the anti-Thy1N model.