Inhibiting miR-155-5p promotes proliferation of human submandibular gland epithelial cells in primary Sjogren's syndrome by negatively regulating the PI3K/AKT signaling pathway via PIK3R1
ZHANG Yuru
WAN Lei
FANG Haoxiang
LI Fangze
WANG Liwen
LI Kefei
YAN Peiwen
JIANG Hui
Abstract：Objective To investigate the mechanism mediating the regulatory effect of miR-155-5p on proliferation of human submandibular gland epithelial cells(HSGECs)in primary Sjogren's syndrome(pSS).Methods Dual luciferase reporter assay was used to verify the targeting relationship between miR-155-5p and the PI3K/AKT pathway.In a HSGEC model of pSS induced by simulation with TRAIL and INF-γ,the effects of miR-155-inhibitor-NC or miR-155 inhibitor on cell viability,cell cycle,apoptosis and proliferation were evaluated using CKK8 assay,flow cytometry and colony formation assay.ELISA and RT-PCR were used to detect the expressions of inflammatory cytokines and miR-155-5p mRNA in the cells;Western blotting was performed to detect the expressions of proteins in the PI3K/AKT signaling pathway.Results Dual luciferase assay showed that miR-155-5p targets the PI3K/AKT pathway via PIK3R1 mRNA.The HSGEC model of pSS showed significantly decreased cell viability,cell clone formation ability and expressions IL-10 and IL-4 and increased cell apoptosis,cell percentage in G2 phase,expressions of TNF-α,IL-6,miR-155-5p and PIK3R1 mRNA,p-PI3K/PI3K ratio,p-Akt/AKT ratio,and PIK3R1 protein expression.Treatment of the cell models with miR-155 inhibitor significantly increased the cell viability,G1 phase cell percentage,colony formation ability,and expressions of IL-10 and IL-4 levels,and obviously reduced cell apoptosis rate,G2 phase cell percentage,expressions of TNF-α,IL-6,miR-155-5p and PIK3R1 mRNA,p-PI3K/PI3K ratio,p-AKT/AKT ratio,and PIK3R1 protein expression.Conclusion In HSGEC model of pSS,inhibition of miR-155-5p can promote cell proliferation and reduced cell apoptosis by targeting PI3K1 mRNA to negatively regulate the overexpression of PI3K/AKT signaling pathway.
Keywords：primary Sjogren's syndromemiR-155-5pPIK3R1PI3K/AKT signaling pathwayinflammatory factors
Publication Date：2025-01-19
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 65-71 )