摘要: Objectives Terminally differentiated stratified squamous epithelial cells play an important role in barrier protection of the skin. The integrity of epidermal cells is maintained by tight regulation of proliferation and differentiation. The aim of this study was to investigate the role of epigenetic regulator H3K4me3 and its demethylase Jarid1b in the control of epithelial cell differentiation. Materials and methods RT-qPCR, Western blotting and IHC were used to detect mRNA and protein levels. We analysed cell proliferation by CCK8 assay and cell migration by wound healing assay. ChIP was used to measure H3K4me3 enrichment. A chamber graft model was established for epidermal development. Results Our studies showed that H3K4me3 was decreased during epidermal differentiation. The H3K4me3 demethylase Jarid1b positively controlled epidermal cell differentiation in vitro and in vivo. Mechanistically, we found that Jarid1b substantially increased the expression of mesenchymal-epithelial transition (MET)-related genes, among which Ovol1 positively regulated differentiation gene expression. In addition, Ovol1 expression was repressed by PI3K-AKT pathway inhibitors and overexpression (O/E) of the PI3K-AKT pathway suppressor Ship1. Knockdown (KD) of Ship1 activated downstream PI3K-AKT pathway and enhanced Ovol1 expression in HaCaT. Importantly, we found that Jarid1b negatively regulated Ship1 expression, but not that of Pten, by directly binding to its promoter to modulate H3K4me3 enrichment. Conclusion Our results identify an essential role of Jarid1b in the regulation of the Ship1/AKT/Ovol1 pathway to promote epithelial cell differentiation.

摘要: Objectives Low back pain becomes a common orthopaedic disease today. It is mainly induced by the degeneration of the intervertebral disc. In this study, we tried to reveal the pathogenesis of the degeneration and the relative therapeutic strategy, which are still elusive. Materials and Methods We collected 15 degenerative intervertebral tissues and five healthy donors. Nucleus pulposus and annulus fibrosus cells were subcultured. miR-640 expression was determined by qPCR. Computer analysis and luciferase reporter assay were used to confirm miR-640 target genes. Immunohistochemical and immunocytochemical staining was used to trace the proinflammatory cytokines and key transductor of signalling pathways. We also used beta-galactosidase staining, flow cytometry, and cell viability assay to monitor the degenerative index. Results miR-640 overexpressed in patients derived degenerative nucleus pulposus tissues and cells. The inflammatory environment promoted miR-640 expression via NF-kappa B signalling pathway. In addition, miR-640 targeted to LRP1 and enhances NF-kappa B signal activity, which built a positive feedback loop. miR-640 inhibited the expression of beta-catenin and EP300, therefore, restrained WNT signal and induced the degeneration in nucleus pulposus cells. miR-640 inhibitor treatment exhibited the effects of anti-inflammation, reverse WNT signalling pathway exhaustion, and remission of degenerative characteristics in vitro. Conclusions miR-640 plays an important role in the degeneration of intervertebral disc and the relative inflammatory microenvironment. It is a promising potential therapeutic target for the low back pain biotherapy.

摘要: Objectives To investigate the functions of miR-223-3p and ITGB3 in pulmonary arterial hypertension (PAH). Materials and Methods Microarray analysis was used to detect differentially expressed genes and microRNAs. In in vitro models, the expressions of miR-223-3p and ITGB3 were detected by qRT-PCR and Western blot. alpha-SMA expression and cell proliferation were analysed by immunofluorescence and MTT assay, respectively. In in vivo models, PAH progressions were determined by measuring the levels of mPAP and RVSP. Lung and myocardial tissues were subjected to HE staining and Masson and Sirius red-saturated carbazotic acid staining to investigate the pathological features. Results The microarray analysis revealed that ITGB3 was upregulated, while hsa-miR-223-3p was downregulated in PAH. After the induction of hypoxia, miR-223-3p was downregulated and ITGB3 was upregulated in PASMCs. Hypoxia induction promoted cell proliferation and inhibited alpha-SMA expression in PASMCs. Both the upregulation of miR-223-3p and the downregulation of ITGB3 attenuated the aberrant proliferation induced by hypoxia conditions. After approximately 4 weeks, the mPAP and RVSP levels of rats injected with MCT were decreased by the overexpression of miR-223-3p or the silencing of ITGB3. The staining results revealed that both miR-223-3p overexpression and ITGB3 knockdown alleviated the pulmonary vascular remodelling and improved the PAH pathological features of rats. Conclusions MiR-223-3p alleviated the progression of PAH by suppressing the expression of ITGB3, a finding which provides novel targets for clinical treatment.

摘要: Objectives The lung-gut axis is known to be involved in the pathogenesis of Staphylococcus aureus pneumonia. However, the underlying mechanisms remain unclear. We examined the role of pulmonary mast cells (MCs) in the regulation of the lung-gut axis during S. aureus pneumonia. Materials and Methods We created a mouse model of S. aureus pneumonia using MC-deficient mice (Kit(W-sh/W-sh)) and examined the level of inflammation, bacterial burden, expression of cathelicidin-related antimicrobial peptide (CRAMP) and composition of the gut microbiota. We further evaluated anti-bacterial immunity by administering bone marrow MCs (BMMCs) or CRAMP into the lungs of Kit(W-sh/W-sh) mice. Results After S. aureus challenge, the MC-deficient mice, compared with wild-type (WT) mice, displayed attenuated lung inflammation, decreased expression of CRAMP, higher bacterial lung load and disturbance of the intestinal microbiota. Adoptive transfer of BMMCs into the lung effectively reconstituted the host defence against S. aureus in Kit(W-sh/W-sh) mice, thus resulting in recovery of S. aureus pneumonia-induced intestinal dysfunction. Similarly, exogenous administration of CRAMP significantly enhanced anti-bacterial immunity in the lungs of MC-deficient mice. Conclusions This study provides evidence for the involvement of MCs in the regulation of the lung-gut axis during S. aureus pneumonia.

摘要: Objectives Thyrotroph embryonic factor (TEF) plays an important role in several different processes in normal human cells; however, its function in malignant cells has not been fully elucidated. Materials and methods The mRNA levels of TEF in 408 bladder cancer (BC) samples from the Cancer Genome Atlas (TCGA) database were analysed in depth. Next, the expression of TEF in 7 BC cell lines was compared to that in normal bladder epithelial cells. The cell count, colony formation and anchorage-independent growth assays as well as a nude mouse xenograft model were utilized to examine the effects of TEF on proliferation and tumorigenesis. Immunofluorescence staining, flow cytometry analysis and treatment with an AKT inhibitor were performed to explore the molecular regulation mechanisms of TEF in BC. Results Analysis of TCGA data indicated that TEF mRNA was decreased in BC samples compared to that in normal bladder epithelial cells and correlated with the poor survival of BC patients. Additional experiments verified that the mRNA and protein expression of TEF were significantly decreased in BC cells compared to that in normal bladder epithelial cells. Upregulation of TEF expression significantly retarded BC cell growth by inhibiting the G1/S transition via regulating AKT/FOXOs signalling. Conclusion Our results suggest that TEF might play an important role in suppressing BC cells proliferation and tumorigenesis.