Abstract: SA-PO080
MCM4 Protects Against Cisplatin-Induced AKI Through Regulating the 53BP1/USP28/p53/p21 Signaling Pathway
Session Information
- AKI: Mechanisms - III
November 05, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Huang, Jing, Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
- Su, Hua, Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
- Zhang, Chun, Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
Background
The minichromosome maintenance (MCM) proteins family exert an essential role in DNA replication and DNA damage repair. Recently, it has reported that MCMs are implicated in various tumors and their expression correlates with the prognosis and recurrence of tumors. However, the role of MCMs in kidney diseases remains unknown. In this study, we aimed to explore the role of MCM4 in the pathogenesis of cisplatin-induced AKI.
Methods
Male C57 mice were intraperitoneally injected with cisplatin for 3 days. In vitro, NRK-52E cells were treated with cisplatin for 12h. Expressions of MCM4, NGAL, cleaved-caspase-3, Bax, p53, p21, 53BP1, USP28 and MAD2B were detected by Western blot and immunohistochemistry. MCM4, 53BP1 and USP28 knockdown were achieved by transfected with siRNA, respectively. Lentivirus transfection was utilized to knock down the expression of MAD2B. Flow cytometry was conducted for cell cycle. CCK-8 assay was conducted to evaluate the cell viability. TUNEL assay was used to detecte the apoptotic cells and ROS detection kit was utilized for ROS detection.
Results
Here, we show that MCM4 is highly expressed in tubules while less in glomeruli, and the expression of MCM4 is upregulated in mice and NRK-52E cells under cisplatin treatment. Silencing of MCM4 inhibits cell proliferation and delays the entry of S phase, and then aggravates cell apoptosis in cisplatin-treated NRK-52E cells. Mechanistically, MCM4 deficiency results in augmented activity of the 53BP1/p53/p21 pathway, which is a key pathway for regulation of cell proliferation and apoptosis. Moreover, based on the previous studies of MAD2B in kidney diseases, we find that MAD2B deficiency inhibits the activity of CDK/Cyclin complex, the phosphorylation of Rb and the transcription level of E2F1, leading to the downregulation of MCM4.
Conclusion
Therefore, our data revealed the protective role of MCM4 in cisplatin-induced acute kidneyr injury. MCM4 was involved in cell proliferation and apoptosis by 53BP1/USP28/p53/p21 signaling pathway under the cisplatin stimulation. On the other hand, MAD2B could influence the the phosphorylation of Rb and the transcription level of E2F1, resulting in the changes in MCM4 proteins levels. In conclusion, MCM4 maybe a potential therapeutic target for the treatment of AKI.