PMC

circRNA-miRNA-mRNA network. The network illustrates 44 molecules, in which diamonds represent mRNA, hexagons represent miRNA, and triangles indicate circRNA. The interactions between them are shown by edges.

Circ_0001859 inhibits the transcription activity of miR-204. Effects of empty vector (EV), circ_0001859, circ_0001859 inhibitor (circ-in), miR-204/211 inhibitor (miR-in), and miRNA inhibitor control (IC) on the luciferase activity of miR-204 in SW982. ^∗ P < 0.05.

MiR-204/211 targeting of ATF2 mRNA. Luciferase reporter analysis of wild-type or mutant ATF2 3′-UTR. MiR-204 was transfected with wild-type or mutant vectors. The values are the mean ± SEM of three replicates. ^∗ P < 0.05.

RA-related molecular signaling pathway. ATF2 is involved in the TLR signaling and TNF pathways. ATF2 regulates transcriptions with the c-Fos protein to regulate the expression of GMCSF, CCL5, IL-6, and IL-1b. Hsa_circ_0001859 can also regulate ATF2 by inhibiting miR-204/211.

Effects of hsa_circ_0001859 on gene expression in SW982 cells. (a) The influence of three different siRNAs against hsa_circ_0001859 was analyzed by quantitative polymerase chain reaction (qPCR), and siRNA-circ0001859-1 (si-1) exhibited the best inhibitory effect. (b) The expression levels of ATF2, c-Jun, c-Fos, TNF, IL-1β, and IL-6 were analyzed by Western blotting. The values are the mean ± SEM of three replicates. P < 0.05. GAPDH was used as a loading control. (c) ATF2, c-Jun, c-Fos, TNF, IL-1β, and IL-6 mRNA expression levels were analyzed after knockdown of hsa_circ_0001859 by siRNA-circ0001859 or cotransfection with siRNA-circ0001859 and the miR-204/211 inhibitor. ^∗ P < 0.05.

Differential expression of genes in RA synovial tissues. (a) The fold value and P value were used to build the volcano plots. The horizontal line corresponds to twice the normal and downward adjustment between the normal and RA samples, and the vertical line shows the P value. The red points and green points in the plot represent the total of 369 DEGs with statistical significance. (b) Hierarchical clustering analysis of DEGs. Each group has four components (greater than two-fold expression difference; P < 0.05). The expression values are expressed in different colors to indicate expression above or below the median expression level for all samples.