POS-079 14-3-3ζ：A PROTECTOR IN CISPLATIN-INDUCED ACUTE KIDNEY INJURY

Abstract

Cisplatin is an anticancer agent marred by acute kidney injury (AKI). Limiting this adverse effect may allow the use of higher doses to improve its efficacy. Basing on our previous study verifying that exosomes from bone marrow mesenchymal stem cells (BMSCs) can promote the repair in cisplatin-induced AKI(AKI), the objective of this study was to identify a candidate that targets to alleviate CKI. We analyzed cisplatin-injured cells and BMSCs-treated cells by using iTRAQ isobaric tags and 2D nano LC-MS/MS.14-3-3 expression in the rat renal tubule epithelial cells (NRK-52E) was evaluated through RT-qPCR and western blotting analyses. 14-3-3ζ overexpression lentiviruses and small interfering RNA (siRNA) transfection were applied to test the its function in CKI. 1. By using the iTRAQ method, we analyzed 3539 proteins, of which 266 showed significantly different expression in BMSCs-treated cells, including 14-3-3. Ingenuity pathway analysis (IPA) showed that the majority of differentially expressed proteins (DEPs) were associated with cell death and survival, cell cycle, cellular movement and development. Further analysis indicated that 14-3-3 had the strongest effect on cell cycle and apoptosis.2. The expression of 14-3-3ζ in NRK-52E was downregulated when administered with cisplatin. Along with this, the cell viability and proliferation level in cisplatin group were dramatically reduced as measured by cck8 and flow cytometry analysis. Besides, the proportion of apoptotic cells increased sharply comparing to control group.3. pro-apoptotic protein cleaved-caspase3 increased and anti-apoptotic protein bcl-2 reduced. Further, Co-IP assays showed the interaction between 14-3-3ζ and β-catenin. Si14-3-3ζ downregulated expression of β-catenin and its nuclear translocation as revealed by western blot and immunofluorescence analysis. Besides, lower β-catenin nuclear translocation induced by knockdown of 14-3-3ζ decreased transcription of cyclin D1.4. Similarly, overexpression of 14-3-3ζ exerted a positively protective effect on NRK-52E cells administered with cisplatin, accompanied by the motivation of β-catenin/cyclinD1 pathway. Together, our findings indicate that 14-3-3ζ interacts with β-catenin, contributing to proliferation and apoptosis through β-catenin/cyclinD1 pathway in CKI, which lead to a previously unrecognized dual function and have potential for treatment of CKI.