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<p>Although RNF13 is known to be dysregulated in models of retinal degeneration, its specific role in retinal pigment epithelium cell function remains unclear. This study aimed to elucidate the RNF13-regulated signalling pathways and identify key molecular targets in vitro. Using lentivirus-mediated RNF13 knockdown in ARPE-19 cells, we observed significant alterations in protein expression using tandem mass tag proteomics and identified 340 differentially expressed proteins (DEPs), among which 80 were downregulated and 260 were upregulated. GO annotation revealed that these DEPs were primarily associated with “membrane” components and linked to “cellular processes” and “metabolic processes” involving “binding” and “catalytic activity”. KEGG pathway analysis revealed significant disruptions in metabolic pathways, particularly the PPAR signalling pathway. Subsequent validation via western blotting, qRT-PCR, and parallel reaction monitoring confirmed that SCD expression is downregulated in RNF13-knockdown cells. Furthermore, siRNA-mediated knockdown of SCD expression resulted in a significant reduction in the Oil Red O-stained area and inhibited cellular proliferation. Impaired lipid metabolism and cell viability induced by RNF13 knockdown were restored by SCD overexpression. These findings suggest that RNF13 may influence lipid metabolism through the PPAR signalling pathway in ARPE-19 cells, with a possible involvement of SCD as a potential mediator in this process.</p>

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Proteomic analysis reveals lipid metabolism disruption and key targets in ARPE-19 cells after RNF13 knockdown

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月10日

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  • Chen Xie,
  • Xin Yu,
  • Liyin Wang

摘要

Although RNF13 is known to be dysregulated in models of retinal degeneration, its specific role in retinal pigment epithelium cell function remains unclear. This study aimed to elucidate the RNF13-regulated signalling pathways and identify key molecular targets in vitro. Using lentivirus-mediated RNF13 knockdown in ARPE-19 cells, we observed significant alterations in protein expression using tandem mass tag proteomics and identified 340 differentially expressed proteins (DEPs), among which 80 were downregulated and 260 were upregulated. GO annotation revealed that these DEPs were primarily associated with “membrane” components and linked to “cellular processes” and “metabolic processes” involving “binding” and “catalytic activity”. KEGG pathway analysis revealed significant disruptions in metabolic pathways, particularly the PPAR signalling pathway. Subsequent validation via western blotting, qRT-PCR, and parallel reaction monitoring confirmed that SCD expression is downregulated in RNF13-knockdown cells. Furthermore, siRNA-mediated knockdown of SCD expression resulted in a significant reduction in the Oil Red O-stained area and inhibited cellular proliferation. Impaired lipid metabolism and cell viability induced by RNF13 knockdown were restored by SCD overexpression. These findings suggest that RNF13 may influence lipid metabolism through the PPAR signalling pathway in ARPE-19 cells, with a possible involvement of SCD as a potential mediator in this process.