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Objective @#To investigate the effect and mechanism of methyltransferase-like 3 (METTL3) on the pro- liferation , migration , and secretion of inflammatory factors by synovial fibroblasts from rheumatoid arthritis (RA) .@*Methods @#The expression of METTL3 in synovial tissue (SF) from 25 patients with rheumatoid arthritis and 25 pa- tients with osteoarthritis was detected by RT-qPCR and immunohistochemistry , respectively . The concentration of RNA m6A was detected by ELISA . RA synovial fibroblasts were isolated and cultured , and divided into NC ( nor- mal control) group , hi-METTL3 (overexpression of METTL3) group , si-METTL3 (knock-down METTL3) group , and STM2457 (METTL3 specific inhibitor) intervention group . Cell proliferation was detected by CCK-8 method . Apoptosis was detected by flow cytometry . And the concentrations of interleukin-6 ( IL-6) , interleukin-17A ( IL- 17A) , receptor activator of nuclear factor-kappa B ligand (RANKL) , and osteoprotegerin (OPG) in the superna- tant of cell culture were detected by ELISA . @*Results @#Compared with synovial tissue of osteoarthritis , the expres- sion of mRNA m6A and METTL3 in synovial tissue of RA significantly increased (P < 0. 05) . After overexpression of METTL3 , the expression of m6A in synovial fibroblasts increased . The proliferation and migration abilities of SF in hi-METTL3 group were significantly improved , and their apoptosis did not change significantly . The secretion of cytokines IL-6 and RANKL of SF in hi-METTL3 group significantly increased , while the OPG significantly de- creased (P < 0. 05) . After interfering with METTL3 expression , the expression of m6A in synovial fibroblasts de- creased . Cell proliferation and migration of SF in siMETTL3 group significantly decreased . The secretion of cyto- kines IL-6 and RANKL significantly decreased , and OPG significantly increased ( P < 0. 05) . After intervention with METTL3 inhibitor STM2457 , the proliferation and migration of synovial fibroblasts were significantly reduced , and the secretion of cytokines IL-6 and RANKL significantly reduced , and OPG significantly increased ( P < 0. 05) . There was no significant difference in the expression of IL-17A among each group . @*Conclusion @#METTL3 may promote the proliferation and migration of RA synovial fibroblasts , enhance the expression of IL-6 and RANKL , and inhibit the expression of OPG through RNA m6A methylation modification .
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Objective To investigate the real-time regulatory effects of IFN-γ, programed death ligand 2(PDL2) and janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway on the adherence, proliferation and migration of human placenta-derived mesenchymal stem cells(hPMSCs) based on a finding that IFN-γ could enhance the expression of PDL2 in hPMSCs through JAK/STAT signaling pathway.Methods hPMSCs were isolated by using enzyme digestion method and then co-cultured with IFN-γ, anti-PDL2 monoclonal antibody (anti-PDL2 McAb) and JAK inhibitor, respectively.Real-time cell analysis (RTCA) was used to detect the dynamic changes in the adherence, proliferation and migration of hPMSCs following various interventions.Results IFN-γ remarkably suppressed hPMSCs proliferation during the period from 40 hours to 80 hours after intervention and also inhibited the non-targeted migration of hPMSCs.However, hPMSCs adherence was not affected by IFN-γ.Co-culturing hPMSCs with anti-PDL2 McAb significantly enhanced hPMSCs adhesion and inhibited their non-targeted migration, but had no significant effect on hPMSCs proliferation.Furthermore, the proliferation of hPMSCs co-cultured with IFN-γ and anti-PDL2 McAb was significantly inhibited as compared with that of anti-PDL2McAb treatment group.The adhesion, migration and proliferation of hPMSCs were significantly inhibited after co-culturing them with JAK inhibitor.Conclusion IFN-γ can remarkably suppress the proliferation and migration of hPMSCs.PDL2 can enhance the migration and inhibit the adhesion of hPMSCs.JAK/STAT signaling pathway is involved in regulating the adhesion, migration and proliferation of hPMSCs.
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Objective To investigate the correlations between the soluble form of B7-H3 ( sB7-H3) and the cytokines of IL-17 and IL-8 in serum samples from patients with primary hepatocellular carcino-ma ( HCC) and to evaluate their clinical values for early diagnosis of HCC.Methods Serum samples were collected from 63 patients with HCC and 50 healthy subjects.The expression of sB7-H3, IL-17 and IL-8 in serum samples were detected by ELISA.Receiver operating characteristic ( ROC) curve was generated to an-alyze the diagnostic values of sB7-H3, IL-17 and IL-8 for hepatoma.The logistic regression model was used to predict the probability of hepatoma by using sB7-H3, IL-17 and IL-8 in combination.Results The levels of sB7-H3, IL-17 and IL-8 in serum samples collected from the patients with HCC were significantly higher than those from healthy subjects.A positive correlation was found between the levels of sB7-H3 and IL-17 in serum samples from patients with HCC.No correlation was found between sB7-H3 and IL-8.A negative cor-relation was found between the levels of IL-17 and IL-8 in serum samples from patients with HCC.ROC curve analysis showed that the area under the curve (AUC) of sB7-H3, IL-17 and IL-8 were 0.832, 0.657 and 0.953, respectively, indicating the statistical significance of them for the diagnosis of HCC.The logistic regression showed that the AUC, diagnostic sensitivity and specificity of the regression model PRE in the pre-diction of HCC were 0.960, 91.30% and 94.29%, respectively, which was much better than using the three indicators alone.Conclusion The levels of sB7-H3 were positively correlated to the levels of IL-17 in serum samples from patient with HCC.The logistic regression model of combination of sB7-H3, IL-17 and IL-8 obtained in this study could be used for early clinical diagnosis of HCC in the future.