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Objective: To analyze the expression of lnc-MTBP-5 in colorectal cancer (CRC), and explore the effect of lnc-MTBP-5 on the invasion of CRC cells and its potential mechanism. Methods: Bioinformatics data from PRJNA218851 and PRJNA376161 data sets were extracted from the Sequence ReadArchive (SRA) database to screen CRC metastasis associated lncRNAs. The Cancer Genome Atlas (TCGA) was used to analyze the expression of lnc-MTBP-5 in CRC tissues and normal tissues, its relationship with the prognosis of patients, and its correlation with metastasis related factors. Quantitative real-time PCR (qPCR) was used to detect the expression of lnc-MTBP-5 in normal intestinal epithelial cells and CRC cells, as well as 53 CRC tissues and para-cancer mucosa. After lnc-MTBP-5 was down-regulated in CRC cells, CCK-8 assay, clone formation and Transwell assay were performed to observe the effect of lnc-MTBP-5 on the proliferation and invasion ability of CRC cells. Results: Lnc-MTBP-5 was associated with CRC metastasis. The expression of lnc-MTBP-5 was significantly increased in 5 CRC cell lines and CRC tissues. Compared with patients with low expression of lnc-MTBP-5, CRC patients with high expression of lnc-MTBP-5 were younger, had higher American Joint Committee on cancer (AJCC) staging, and were prone to metastasis. Lnc-MTBP-5 was positively correlated with CRC metastasis associated in colon cancer 1 (MACC1), mesenchymal to epithelial transition factor (MET) and cadherin-associated protein. After lnc-MTBP-5 was down-regulated, the invasion ability of CRC cells decreased. Conclusion: Lnc-MTBP-5 is up-regulated in CRC cell lines and CRC tissues, and it is negatively correlated with the prognosis of patients. Lnc-MTBP-5 can promote the invasion ability of CRC cells, which may be related to MACC1-HGF (hepatocyte growth factor)/MET pathway.
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Objective To investigate the immunosup-pressive activity of benzoxazole derivative PO-291 in inhibiting human activated T cell proliferation and function. Methods Human T cells were isolated and purified by the immunomagnetic microbeads and acti-vated by anti-CD3/anti-CD28 mAbs or alloantigen. Cell proliferation, the expression of CD25 and CD69, cell cycle and apoptosis were measured by flow cytome-try. Secretion levels, including IL-2, IL-4, IL-6, IL-10, IL-17 and IFN-γ were determined by ELISA. The expression and phosphorylation of STAT5 and p70S6K of activated T cells were detected by Western blot. Re-sults PO-291 significantly inhibited human T cell proliferation with anti-CD3/anti-CD28 mAbs or alloan-tigen stimulation without obvious cytotoxicity. PO-291 did not affect CD25, CD69 and IL-2 expression, but induced T cell cycle arrest in G0/G1 phase. PO-291 significantly inhibited IL-17, IFN-γ and IL-6 expres-sion, but not IL-2, IL-4 and IL-10. PO-291 did not affect STAT5 and p70S6K expression, but inhibited STAT5 phosphorylation and enhanced p70S6K phos-phorylation. Conclusions PO-291 inhibits human ac-tivated T cell proliferation by affecting the JAK3/STAT5 pathway. PO-291 represents a potential lead compound for the design and development of new im-munosuppressive drugs for the treatment of organ trans-plantation and autoimmune diseases.
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Objective:To investigate the effects of microgroove surface morphology on the adhesion and proliferation of the human gin-gival fibroblasts(HGFs).Methods:Microgroove surfaces of titanium were fabricated by photolithography with parallel grooves of 15,30 or 60 μm in width and 5 μm or 10 μm in depth.The groups of the samples were denoted as T15 /5,T15 /10,T30 /5,T30 /10,T60 /5 and T60 /10.Smooth titanium surface(T0)was used as the control.The surface topography was observed by enviroment SEM(ES-EM).HGFs were cultured on the topographically modified surfaces.Morphology was observed by SEM.Cell proliferation was examined by CCK-8 kit.Results:HGFs on the microgroove surfaces had “contact guidance”parallel to the microgrooves,whereas the cells on T0 were oriented randomly.Cell proliferation was promoted and kept for longer period on T60 /10 surface.Conclusion:Surfaces of mi-crogrooves with increasing groove width and depth may achieve “contact guidance”for HGFs and promote cell proliferation.
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Objective To investigate the optimal condition of lentivirus,which was recombined with marker gene of enhanced green fluorescent protein (Lentivirus-EGFP) transfect human umbilical cord wharton’s jelly-derived mesenchy-mal stem cells (HUWMSCs) and the effect of transfection on the proliferation in HUWMSCs. Methods HUWMSCs were transfected with EGFP by lentivirus vector in vitro via different multiplicity of transfection (MOI) in four different transfec-tion methods (A, B, C and D). The fluorescence expression and the transfection efficiency in different methods were analyzed by both fluorescent microscope and flow cytometry. The proliferation rates of infected HUWMSCs was evaluated by MTT method. Results The transfection efficiency was 10.6%-87.3%after 4 days in all experimental groups, which showed the dose-effect relationship with MOI. Polybrene (5 mg/L) could significantly increase the transfection efficiency (P＜0.05). Re-sults of MTT assay showed that there were significant differences in the proliferation rates of infected HUWMSCs between different transfection methods (P ＜ 0.001). There was better cell proliferation in method A (MOI=10) group and method B (MOI=10) group than that of other groups. Conclusion Method B (MOI=10) is the optimal transfection method in this exper-iment. HUWMSCs could be transfected by lentivirus-EGFP with high efficiency and could stably express transfected gene within 2 weeks, which is a safe and effective gene transfer vector.