摘要: Objective: Aberrant activation of Wnt/beta-catenin signalling contributes significantly to the development of human colorectal cancers and beta-catenin is the key signalling molecule transducing canonical Wnt/beta-catenin signalling. Therefore, beta-catenin is a promising therapeutic target for cancer treatment. This study demonstrates that the oncogenic IKKe kinase phosphorylates beta-catenin to restrain its hyper activation, therefore promoting colorectal cancer (CRC) cell proliferation. Materials and methods: IKKe and beta-catenin expression levels in human colorectal cancer tissues and cell lines were analysed by immunohistochemical staining and Western blotting. The regulation of IKKe on Wnt/beta-catenin signalling pathway was studied by reporter assay and real-ime PCR analysis in the context of IKKe stably knocking down. Co-immunoprecipitation was conducted to monitor the interaction between IKKe and beta-catenin. Kinase assay was performed to measure beta-catenin post-translational modifications induced by IKKe. Results: Oncogenic IKKe kinase is required for the proliferation of colorectal cancer cells. Mechanistically, inhibition of IKKe results in beta-catenin hyper activation and thwarts CRC cell proliferation. Furthermore, IKKe phosphorylates beta-catenin and inhibits the activation of beta-catenin signalling. Conclusion: Our study suggests that IKKe is a potential target to combat CRC induced by aberrant Wnt/beta-catenin signalling.

摘要: Objectives: Schwann cells (SCs) are the principal glial cells in peripheral nerve system, involved in neuropathies with great regenerative potential. Dental pulp cells have been reported to maintain neurogenic potential. In contrast, the regulatory role of SCs on human dental pulp cells (hDPCs) development remains undefined. Materials and methods: SC secretion and SC-derived extracellular vesicles (EVs) were collected and used to treat hDPCs; and proliferation and multiple differentiation of hDPCs were detected after EVs treatments. Finally, we analysed the proteomes of SC-EVs and SCs through mass spectrum. Results: In this study, we found SC secretion showed a predominantly regulatory role on the development of hDPCs. Further, we identified EVs from SC secretion with similar function as SC secretion in regulating hDPCs proliferation and multipotency. And expression of transcription factor Oct4 was upregulated after treatment of both SC secretion and EVs, as well as Sox2 and Nanog. We detected abundant enrichment of Oct4 in EVs, which might be responsible for the upregulation of stem cell-related genes in hDPCs. Through proteome and western blot analysis, we found enriched TGF beta s in EVs, indicating that accelerated hDPCs proliferation may be mediated by activated TGF beta-Samd and TGF beta-MAPK signalling. Conclusions: In summary, our study sheds light on critical regulatory ability of SC-derived EVs on hDPCs proliferation and multipotency, suggesting great implications for seeding cells used in tissue engineering.

摘要: Objectives: The aim of this study was to investigate the role of insulin-like growth factor-1 (IGF-1) and crosstalk between endothelial cells (ECs) and adipose-derived stem cells (ASCs) in the process of angiogenesis. Methods: A three-dimensional collagen gel used to culture mouse ASCs and mouse ECs in vitro was established. The effects of angiogenesis after exposure to IGF-1 were observed by confocal laser scanning microscopy. Western blotting and qPCR were performed to elucidate the underlying mechanisms. Results: IGF-1 treatment promoted the formation of vessel-like structures and the recruitment of ASCs in the three-dimensional collagen gel. The angiogenic genes and proteins in ECs were up-regulated by IGF-1 and in co-culture. Similar changes in the genes and in the proteins were detected in ASCs after exposure to IGF-1 and co-culture. p-Akt expression levels were high in ECs and ASCs after exposure to IGF-1 and co-culture. Conclusions: IGF-1 and co-culture between cells facilitate the process of angiogenesis via the PI3-kinase/Akt signalling pathway. In ECs, IGF-1 stimulates the expression of angiogenesis-related growth factors with the activation of the PI3-kinase/Akt signalling pathway. Co-cultured ECs exposed to excess VEGF-A and other angiogenesis-related growth factors para-secreted from ASCs exhibit high expression of angiogenesis-related genes and proteins. In ASCs, IGF-1 induces the recruitment and function of ASCs by up-regulating the expression of PDGFB, MMPs and alpha-SMA. Crosstalk with ECs further facilitates changes in ASCs.

摘要: Objective: To study the effect on endometrial and endometriotic cells after co-culture with macrophages, using clonogenic, invasion and self-renewal assays. Materials and methods: Peripheral blood samples, endometrium and endometriotic tissues were collected. Autologous macrophages were co-cultured with endometrial and endometriotic cells. The number of colony-forming units (CFU), invasiveness and self-renewal activity after co-culture with macrophages were determined. The cytokine level of colony-stimulating factor-1 (CSF-1) from macrophages with and without endometriosis was compared. Results: Co-culture with macrophages significantly increased the clonogenic and invasion ability of endometriotic stromal cells in vitro. Colony-stimulating factor-1 (CSF-1) was up-regulated in endometriotic macrophages conditioned medium when compared to those without the disease. Conclusions: These data suggest that macrophages may increase the proliferation and invasion activity of stromal clonogenic cells in women with endometriosis.

摘要: Objectives: Serum amyloid A (SAA), an acute phase protein, is highly expressed in psoriatic lesions but its function is not fully understood. The aim of this study was to explore its role in activation of keratinocytes. Materials and methods: Real-time PCR and immunofluorescence were performed to examine SAA expression in imiquimod (IMQ)-induced psoriasis-like mice. In vivo function of SAA was examined by treating psoriasis-like mice with SAA neutralising antibody. Cell viability was monitored using the CCK-8 assay. Real-time PCR was performed to determine expression of genes associated with differentiation and inflammation. Ki67(+) percentage and immunological markers were analysed by flow cytometry. Involvement of formyl peptide receptor-like 1 (FPRL1) in SAA signal transduction was determined by RNA interference. Binding of SAA and FPRL1 was examined by co-immunoprecipitaion. Western blotting was conducted to assess phosphorylation of downstream signalling molecules. Results: SAA was highly expressed in skin lesions of IMQ-treated psoriasis-like mice and neutralising SAA attenuated epidermal hyperplasia and inflammation. SAA in vitro promoted keratinocyte proliferation and expression of immunological mediators, while inhibiting differentiation. Effects of SAA on keratinocyte proliferation and inflammation were mediated by FPRL1, as well as activation of the PI3K/Akt pathway. Conclusions: These observations indicate that SAA/FPRL1 contributed to pathogenesis of psoriasis by promoting keratinocyte proliferation and inflammation, thus providing a potential therapeutic target for disease therapy.