ObjectiveTo explore the mechanism of circ_AFF2 regulating the proliferation, migration, and invasion of rheumatoid arthritis (rheumatoid arthritis, RA) synovial fibroblasts by regulating the phosphatidylinositol 3-kinase (PI3K)/serine/protein kinase B (AKT) pathway.
MethodsqRT-PCR was used to detect the relative expression of circ_AFF2 in healthy control synovial tissue and RA synovial tissue.By culturing RA synovial fibroblast MH7A cells in vitro, the cells were divided into si-NC group (transfected with si-NC), si-circ_AFF2 group (transfected with si-circ_AFF2), and si-circ_AFF2+IGF-1 Group (after transfection of si-circ_AFF2, treated with 50 ng/mL PI3K/AKT pathway activator IGF-1).Then qRT-PCR was used to detect the relative expression of circ_AFF2 in each group of cells; MTT assay was used to detect cell viability in each group; Western blotting was used to detect the expression of Ki-67, PCNA, MMP-2, E-cadherin, N-cadherin, and PI3K/AKT pathway-related proteins in each group of cells; Transwell assay detects cell migration and invasion ability of each group.
ResultsCompared with healthy control synovial group, the relative expression of circ_AFF2 was significantly increased in RA synovial tissue (P < 0.01).Compared with the si-NC group, the relative expression of circ_AFF2, cell viability, Ki-67 and PCNA protein expressions in the si-circ_AFF2 group were significantly decreased (P < 0.01).Compared with the si-NC group, the number of migrating cells, the number of invasive cells, the expression of MMP-2 and N-cadherin protein in the si-circ_AFF2 group were significantly decreased (P < 0.01), and the protein expression of E-cadherin was significantly increased (P < 0.01).The protein expressions of p-PI3K and p-AKT in the three groups were significantly decreased (P < 0.01), and the protein expressions of p-PI3K and p-AKT in the si-circ_AFF2 group and the si-circ_AFF2+IGF-1 group were lower than those in the si-NC group (P < 0.05), while there was no change in the expression of PI3K and AKT among three groups (P>0.05).Compared with si-circ_AFF2 group, the relative expression of circ_AFF2, cell viability, number of migrating cells, number of invasive cells, Ki-67, PCNA, MMP-2 and N-cadherin protein expressions in si-circ_AFF2+IGF-1 group were significantly increased (P < 0.01), and E-cadherin protein expression was significantly decreased (P < 0.01).
ConclusionsCirc_AFF2 can promote the proliferation, migration and invasion of RA synovial fibroblasts by activating the PI3K/AKT pathway.
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