the indicated group Considering that circ_0007059 restored the IFN-impaired proliferation of RMCs and HEK293 cells, we hypothesized that circ_0007059 may also inhibit cell apoptosis in IFN-induced RMCs and HEK293 cells. cytometry were used to assess cell proliferation, viability, and apoptosis of cells. Bioinformatics analysis and dual luciferase reporter assay detect the interaction of circ_0007059, miRNA-1278, and SHP-1. Glomerulonephritis was performed in a mouse model by administration of IFN-expressing adenovirus. IHC staining showed the pathogenic changes. Results In the present study, the expression of circ_0007059 in type I interferon (IFN)-treated renal mesangial cells (RMCs), lupus nephritis (LN) specimens, and HEK293 cells was downregulated compared with that in normal healthy samples and untreated cells. Circ_0007059 overexpression resulted in increased cell proliferation, cell viability, apoptosis, and inflammation-associated factors (CXCL10, IFIT1, ISG15, and MX1) in RMCs and HEK293 cells. In addition, circ_0007059 overexpression significantly restored cell Brequinar proliferation and viability and inhibited IFN-induced apoptosis. Further, the increased expression resulted in reduced inflammation and the downregulation of CXCL10, IFIT1, ISG15, and MX1 in RMCs and HEK293 cells. Circ_0007059 serves as a sponge for miR-1278 so that the latter can target the 3-untranslated region of tests, Brequinar and P? ?0.05 was considered statistically significant. Results Circ_0007059 expression is downregulated in LN specimens and IFN-treated RMCs To study the role of circRNAs in the progression of LN, abnormal circRNA expression was evaluated in renal biopsy samples from patients with SLE using microarray analysis. The expression of several miRNAs in the kidneys of patients with LN was significantly lower than that of the control group, particularly for circ_0007059 (Fig.?1A). To confirm Rabbit polyclonal to ABCA6 these results, RT-qPCR analysis was performed to measure circ_0007059 expression in samples from patients with SLE (n?=?30) and normal samples (n?=?10). The results indicated that the level of circ_0007059 in LN samples was lower than that of healthy controls (Fig.?1B). Because type I IFN is essential for LN in humans and mice, we treated RMCs and HEK293 cells with IFN to induce the LN phenotype in these models (Wolf et al. 2018). Of note, IFN treatment reduced the expression of circ_0007059 (Fig.?1C, D). Hence, our results suggest that circ_0007059 is involved in LN progression. Open in a separate window Fig. 1 Expression of circ_0007059 in kidneys, IFN-induced RMCs and HEK293 cells, and patients with lupus nephritis (LN). A microarray analysis revealed differentially expressed genes between kidney samples from patients with LN (n?=?7) and normal healthy tissue (n?=?7) (biological replicates, 7; technical replicates, 1; repeat time, 3). B the expression of circ_0007059 in renal biopsy samples from patients with LN (n?=?30) and surrounding normal tissue samples (n?=?10) by RT-qPCR (biological replicates, as indicated; technical replicates, 3; repeat time, 3). C, D treatment of RMCs and HEK293 cells with IFN (1,000 units/mL) for 24?h. RT-qPCR was used to measure the expression of circ_0007059 (biological replicates, 3; technical replicates, 2; repeat time, 3). Results are showed as the mean??SEM for biological replicates. *P? ?0.05 vs. the indicated group Circ_0007059 overexpression increases viability and suppresses apoptosis and IFN signaling in RMCs and Brequinar HEK293 cells To analyze the effect of circ_0007059 on the viability of IFN-treated RMCs and HEK293 cells, we transfected cells with a circ_0007059-overexpressing vector or control vector. RT-qPCR data confirmed that the transfection of circ_0007059 resulted in a significant increase in circ_0007059 expression levels (Fig.?2A, B). The CCK-8 assay revealed that RMC and HEK293 cell proliferation were noticeably inhibited at 24, 48 and 72?h after IFN induction, but circ_0007059 expression completely restored proliferation (Fig.?2C, D). Furthermore, as determined by the MTT assay, circ_0007059 overexpression led to a noticeable restoration in cell viability, which was suppressed by IFN-treated RMCs and HEK293 cells (Fig.?2E, F). These results suggested a beneficial role of circ_0007059 on the viability of IFN-treated RMCs and Brequinar HEK293 cells. Open in a separate window Fig. 2 Influence of circ_0007059 overexpression on the cell viability of IFN-induced RMC and HEK293 cells. RMC and HEK293 cells were transfected with a circ_0007059-overexpressing vector or NC vector for 24?h, followed by treatment with 1,000 units/mL IFN for 24?h. A, B Expression level of circ_0007059 in RMC and HEK293 cells was detected by RT-qPCR.
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