Effects of miR-27a on the biological behavior of rheumatoid arthritis syn-ovial cells via TLR4/NF-κB signaling pathway
Aim:To explore the effects of miR-27a on the biological behavior of rheumatoid arthritis synovial cell via Toll-likereceptor(TLR)4/NF-κBsignalingpathway.Methods:Atotalof30 RApatients(RAgroup)whounderwentknee replacement surgery and 18 patients(control group)who underwent emergency trauma amputation during the same period were selected and the expression of miR-27a in the synovial tissue was detected using qRT-PCR.RA fibroblast like synovial cells MH7A were divided into 4 groups:blank control group,TNF-α group,TNF-α+miR-NC group and TNF-α+miR-27a mimic group.qRT-PCR was used to detect the expression level of miR-27a in the 4 groups,CCK-8 was used to detect cell proliferation,cell clone formation assay was used to detect cell clone formation ability.Transwell assay was used to detect cell invasion and migration ability,and Annexin Ⅴ/PI double staining was used to detect cell apoptosis.The dual luciferase assay was used to detect the targeting relationship between TLR4 and miR-27a.Western blot was used to detect the expres-sions of TLR4,NF-κB,p-TLR4 and p-NF-κB proteins in MH7A cells.Results:The expression level of miR-27a in synovial tissue of the control group and RA group was(1.00±0.08)and(0.36±0.05),respectively,and it was lower in RA group than that in the control group(P<0.001).Compared with the blank control group,the expression level of miR-27a in the TNF-α group and TNF-α+miR-NC group decreased,which was increased in the TNF-α+miR-27a mimic group;com-pared with the TNF-α and TNF-α+miR-NC group,the expression of miR-27a in the TNF-α+miR-27a mimic group in-creased(P<0.05).Compared with the blank control group,the proliferation,clone formation,invasion,and migration abili-ties of MH7A cells in the TNF-α group and TNF-α+miR-NC group increased,while the apoptosis rate was decreased;com-pared with the TNF-α group and TNF-α+miR-NC group,the proliferation,clone formation,invasion,and migration abilities of MH7A cells in the TNF-α+miR-27a mimic group decreased,while the apoptosis rate increased(P<0.05).TLR4 was a targeted gene for miR-27a.Compared with the blank control group,the p-TLR4/TLR4 and p-NF-κB/NF-κB of MH7A cells in the TNF-α group and TNF-α+miR-NC group increased;compared with the TNF-α group and TNF-α+miR-NC group,the p-TLR4/TLR4 and p-NF-κB/NF-κB in the TNF-α+miR-27a mimic group decreased(P<0.05).Conclusion:miR-27a may reduce the proliferation,invasion,and migration of RA fibroblast like synovial cells,and promote apoptosis via inhibi-ting TLR4/NF-κB signaling pathway.
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