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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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Background and purpose:Epithelial ovarian cancer has the highest mortality rate of gynecologic cancers and overall survival rates have improved little in the last 20 to 30 years. CXXC ifnger protein 5 (CXXC5) plays an important role in AML (acute myeloid leukemia) and MDS (myelodysplasia). However, little is known about its clinical signiifcance and biological function in epithelial ovarian cancer. This study aimed to investigate the expression of the CXXC5 in ovarian cancer and the effect of the CXXC5 on ES-2 cell proliferation. Methods:①The alteration of CXXC5 in cancer genomics data of TCGA (Cancer Genome Atlas) was analyzed.②The CXXC5 protein in the tissue chips was detected containing 37 benign ovarian cyst and 173 malignant tumor samples. The relationship between the expression of the CXXC5 with the clinicopathological features of patients with ovarian cancer was analyzed by SPSS software;③The cells with the highest CXXC5 expression quantity from 5 ovarian cancer cells were selected by re-al-time quantitative PCR (qRT-PCR) and Western blot.④ES-2 cells with shRNA stable transfection were construted us-ing the strategy of lentivirus infection and analyzed cell proliferation by cell counting kit-8(CCK8). Results:①Through the TCGA database, CXXC5 ampliifcation was found in 7 of 563 cases.②The CXXC5 expression in ovarian malignant carcinoma (39.3%) was higher than that in benign ovarian cyst (13.5%, P=0.003), the histologic type was highly asso-ciated with CXXC5 (43%in serous, 22.9%in mucinous, 23.5%in endometrioid, 67%in clear cell, P=0.014) and there was a signiifcant correlation between CXXC5 and lymph node metastasis (positive vs negative, P=0.022).③The ES-2 cells with shRNA stable transfection had a growth disadvantage (P<0.05). Conclusion:The CXXC5 gene might have an advantage in proliferation of epithelial ovarian carcinoma and be expected to become the biomarker of poor prognosis.
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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 effects of the 2,3,4′,5-Tetrahydroxystilbene -2-O-beta-D-glucoside on the prolif-eration of PASMCs induced by hypoxia , in orde to search new drugs for the treatment and prevention of hypoxic pulmonary vascular remod -eling.Methods 3%O2 hypoxia was used to induced the proliferation of PASMCs .After hypoxic and TSG treatment for 24h, cell growth was determined by cell counting kit -8 ( CCK-8 ) , cell cycle was analysed by flow cytometry , the mRNA expression of HIF -1αwas measured by quantitative real -time PCR, and the reactive oxygen species ( ROS) production was determined by the fluorescence micro-plate reader .Results TSG can block the proliferation of PASMCs through G 0/G1 to S phase of the cell cycle arrest without cell cytotoxic-ity.Further experiments showed that TSG blocking the proliferation of PASMCs was associated suppression the mRNA expression of HIF -1αand the production of intracellular ROS in hypoxia -stimulated-PASMCs.Conclusion TSG can inhibit the proliferation of pulmo-nary artery smooth muscle cells induced by hypoxia through suppression the mRNA expression of HIF -1αand the production of intracel-lular ROS.
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Micro RNA (miRNA)were small RNAs encoded by the non-coding RNA genes,which functioned through degrada-tion of mRNA or inhibition of mRNA translation.The miRNA was involved in numerous biological processes,including cell proliferation,apoptosis,development and differentiation.And ectopic expression of miRNA could result in diseases,such as neoplastic hematologic disorder and solid tumor.miR-1 06b-25 cluster contained miR-1 06b,miR-93 and miR-25.These miR-NAs were correlated with the development of tumor.Therefore, we reviewed recent articles on the relationship of miR-1 06b-25 cluster with tumor.
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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.
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Rabbit's brain phospholipids (RBP) 100~200 mg ?kg-1 ip or sc showed anti-inflammatory action on four kind of acute inflammatory animal models. Cephalin and Lecithin were the effective component of antiin-flammatory action on the acute inflammatory animal model. But RBP did not show antiinflam-matory action on the chronic proliferative inflammatory model. RBP increased blood carbon particle clearance, perfaneal macrophage phagocytosis and enhanced lymphocyte proliferation,hemolysin antibody in normal and immunode-pression animal. RBP increased the cGMP content in the liver tissue of immunodepression animal, 0. 1~1 mg ?L-1 RBP promoted proliferation but 100 mg ?L-1 RBP inhibited proliferation induced by PHA in vitro on human lymphocyte.