Effects of human cyclooxygenase-2 gene silencing on synovial cells of rheumatoid arthritis mediated by lentivirus.

Abstract

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-1β (IL-1β) and tumour necrosis factor alpha (TNF-α) 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-1β and TNF-α, 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.

• Authors: Ping Leng, Dawei Li, Yi Sun, Yingzhen Wang, Haining Zhang
• Journal: Artificial cells, nanomedicine, and biotechnology (Artif Cells Nanomed Biotechnol) Vol. 46 Issue sup3 Pg. S274-S280 ( 2018) ISSN: 2169-141X [Electronic] England
• PMID: 30314410 (Publication Type: Journal Article)

Chemical References

• IL1B protein, human
• Interleukin-1beta
• RNA, Messenger
• RNA, Small Interfering
• Tumor Necrosis Factor-alpha
• VEGFA protein, human
• Vascular Endothelial Growth Factor A
• Cyclooxygenase 2
• PTGS2 protein, human
• Dinoprostone

Topics

• Arthritis, Rheumatoid (genetics, metabolism, pathology, therapy)
• Cyclooxygenase 2 (biosynthesis, genetics)
• Dinoprostone (biosynthesis, genetics)
• Female
• Gene Expression Regulation, Enzymologic
• Gene Silencing
• Humans
• Interleukin-1beta (biosynthesis, genetics)
• Lentivirus
• Male
• RNA, Messenger (biosynthesis, genetics)
• RNA, Small Interfering (biosynthesis, genetics)
• Synovial Fluid (metabolism)
• Tumor Necrosis Factor-alpha (biosynthesis, genetics)
• Vascular Endothelial Growth Factor A (biosynthesis, genetics)