ABSTRACT
Objective:To study whether autoimmune regulator (Aire) affects macrophage polarization. Methods: The mouse mononuclear macrophage cell line RAW264. 7 cells, stable expressing GFP-Aire protein RAW264. 7 cells ( A33-3 ) and stable expressing GFP protein RAW264. 7 cells (C1-6) were stimulated with LPS,IL-4 and LPS combined with immune complex respectively to make the macrophage polarize to M1 (LPS),M2a (IL-4) and M2b (LPS with immune complex). To investigate the effects of Aire on the various types of macrophages polarization,the M1 related molecules (IL-1α,iNOS and IL-6),M2a related molecules (Arg-1) and M2b related molecule (IL-10) were detected by Real-time PCR. Results:The expression of IL-1α,IL-6 and inducible nitric oxide synthase ( iNOS) ,the products of M1 macrophages, were significantly upregulated in RAW264. 7 cells treated with LPS at 0. 5 μg/ml. The expression of Arg-1 and IL-10 mRNA in RAW264. 7 cells increased in a dose-dependent manner after stimulation with IL-4 or LPS combined with immune complexes, respectively. M1 macrophage-related marker iNOS and IL-1 increased, whereas IL-6 levels decreased in A33-3 cells compared with C1-6 cells after treatment with LPS. The expression of Arg1 and IL-10 were downregulated in A33-3 cells compared with C1-6 cells after IL-4 and LPS combined with immune complexes stimulation,respectively. Conclusion:Aire may promote M1 macrophage polarization while inhibit M2a and M2b macrophage polarization.
ABSTRACT
Objective:To investigate the effect and mechanism of transcription factor Foxp 3 on the proliferation and cell cycle of human lung adenocarcinoma cell line A 549.Methods:We knocked down the expression of Foxp 3 using siRNA.Foxp3 inhibition was detected by RT-PCR.Cell proliferation was detected by MTT.Cell cycle of A549 cells were detected by flow cytometry after the transfection of siRNA.Cell cycle-related checkpoint genes were filtered by RT-PCR.The regulation of Foxp3 on cell cycle-related checkpoint genes were detected by immunofluorescence and dual -luciferase reporter assay system.Results: The proliferation of A549 cells were inhibited after silencing Foxp3,and A549 cells were arrested in G0/G1 cycle.G1/S cycle checkpoint gene CCND1 was down regulated.Mechanism research show that Foxp 3 can regulate the expression of CCND 1 directly.Conclusion: Foxp3 can promote the proliferation of A549 cell line by up regulating G 1/S cycle checkpoint gene CCND 1.This provides a new target for the therapeutic targets of lung adenocarcinoma.