CircZNF644 aggravates lipopolysaccharide-induced HK-2 cell impairment via the miR-140-5p/MLKL axis

Area of Science:

• Molecular Biology
• Cell Biology
• Biochemistry

Background:

• Circular RNAs (circRNAs) are implicated in human diseases, including acute kidney injury (AKI).
• Understanding the specific roles of circRNAs in AKI pathogenesis is crucial for developing targeted therapies.
• circRNA zinc finger protein 644 (circZNF644) has emerged as a potential factor in AKI progression.

Purpose of the Study:

• To investigate the functional role of circZNF644 in an in vitro model of AKI.
• To elucidate the molecular mechanism underlying circZNF644's effects on kidney cells stimulated with lipopolysaccharide (LPS).
• To explore circZNF644 as a potential therapeutic target for AKI.

Main Methods:

• Quantitative real-time PCR (qRT-PCR) for gene expression analysis.
• Cell viability, proliferation, cell cycle, and apoptosis assays (CCK-8, EdU, flow cytometry).
• Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays to determine molecular interactions.

Main Results:

• circZNF644 expression was upregulated in LPS-treated HK-2 cells, an AKI model.
• circZNF644 knockdown reversed LPS-induced inhibition of cell proliferation and cell cycle, and reduced apoptosis and inflammation.
• circZNF644 directly targeted miR-140-5p, which in turn targeted mixed lineage kinase domain like pseudokinase (MLKL); circZNF644 knockdown protected cells via this pathway.

Conclusions:

• circZNF644 knockdown protects HK-2 cells from LPS-induced injury by modulating the miR-140-5p/MLKL axis.
• circZNF644 plays a detrimental role in AKI pathogenesis.
• circZNF644 represents a promising therapeutic target for acute kidney injury.