Effects of human cyclooxygenase-2 gene silencing on synovial cells of rheumatoid arthritis mediated by lentivirus.
ARTIFICIAL CELLS NANOMEDICINE AND BIOTECHNOLOGY(2018)
摘要
The aim of the study is to screen the effective shRNA sequence which can silence the human COX-2 expression level in synovial cells of rheumatoid arthritis (RA) patient transfected by the lentivirus. Four pairs of hCOX-2 shRNA were designed and inserted into lentivirus to form pGPHI/GFP/Neo-shRNA vector. The reconstructed virus was transfected into synovial cells derived from RA patients, and then the expression level of hCOX-2 mRNA and the protein of the inflammatory factors including prostaglandin E2 (PGE2), vascular endothelial growth factor (VEGF), interleukin-1 beta (IL-1 beta) and tumour necrosis factor alpha (TNF-alpha) in the supernatants were examined with real-time PCR and ELISA, respectively. There was no obvious negative influence on cell growth and morphology after hCOX-2 shRNA gene transfection mediated by lentivirus. The hCOX-2 mRNA expression level, as well as the concentration of PGE2, VEGF, IL-1 beta and TNF-alpha, decreased significantly (p<.05). RNAi mediated by lentivirus can significantly inhibit hCOX-2 mRNA expression level in synovial cells of RA patients, so as to reduce the expression of inflammatory cytokines.
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关键词
Rheumatoid arthritis,RNA interference,cyclooxygenase-2,inflammatory cytokines,lentivirus,synovial cells
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