2023, Vol. 27 ›› Issue (5): 695-700

Screening and verification of key genes of necroptosis in osteoarthritis

Yuan Changshen1, Guan Yanbing2, Li Zhe2, Rong Weiming2, Liao Shuning2, Chen Lewei2, Mei Qijie1, Duan Kan1   

Abstract: BACKGROUND: Osteoarthritis is closely related to chronic and low-grade inflammation. Necroptosis is a new cell death pathway with extensive inflammatory characteristics, which can affect the progression of osteoarthritis, but the related targets of necroptosis in osteoarthritis have not been found.
OBJECTIVE: To explore the key genes of necroptosis in osteoarthritis based on bioinformatics and verify them with animal experiments, so as to provide new targets for the prevention and treatment of osteoarthritis from the perspective of necroptosis.
METHODS: Osteoarthritis- and necroptosis-related genes were screened from GEO database and GeneCard database respectively, and then the intersection of them was taken to obtain osteoarthritic necroptosis genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were further carried out to construct a protein-protein interaction network. The key genes of osteoarthritic necroptosis (Hub genes) were identified through five calculation methods using Cytoscape software. Finally, male   Sprague-Dawley rats were selected to metaphase models of osteoarthritis. The expression of Hub genes in the metaphase models of osteoarthritis of male Sprague-Dawley rats detected by RT-PCR.
RESULTS AND CONCLUSION: A total of 38 osteoarthritic necroptosis genes were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that they were significantly enriched in leukocyte activity, T cell receptor and interleukin-17 signal pathway. Five Hub genes, CYBB, FADD, IL1B, MYC and EGFR, were screened by the five different algorithms of Cytoscape. RT-PCR showed that EGFR and IL1B were highly expressed in the metaphase models of osteoarthritis of male   Sprague-Dawley rats (P < 0.05), while CYBB and MYC were lowly expressed (P < 0.05), but FADD had no significant changes (P > 0.05). To conclude, CYBB, IL1B, MYC, and EGFR, as the key genes of osteoarthritic necroptosis, are expected to become new targets for the prevention and treatment of osteoarthritis.
Key words: osteoarthritis, necroptosis, key gene, bioinformatics, experiment validation