ABSTRACT
BACKGROUND: Mesenchymal stem cells (MSCs) are an important component of the in vivo microenvironment and act on multiple biological behaviors of tumor cells. The potential clinical value of MSCs has become an issue of concern in recent years.OBJECTIVE: To investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with umbilical cord-derived MSCs (UC-MSCs) using cDNA microarray.METHODS: In vitro co-culture system was constructed, and then cellular proliferation, apoptosis and differentiation status of NB4 cells treated with UC-MSCs were evaluated. Two cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without UC-MSCs. The probes were labeled with fluorescence dyes individually, hybridized with cDNA microarray, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in two gene expression profiles.RESULTS AND CONCLUSION: UC-MSCs promoted the proliferation and differentiation, while reduced the apoptosis of NB4 cells. The analysis of gene expression profiles indicated that after co-culture with UC-MSCs, 530 genes were up-regulated and 53 genes were down-regulated. Accordingly, specific gene function and pathway signaling related were also regulated to some extent. Overall, UC-MSCs influence can major biological behaviors of NB4 cells by changing expression of a large amount of genes, gene-related function and multiple intracellular signaling pathways.
ABSTRACT
ObjectiveToll-like receptors (TLRs) play important role in the progression and tumor immunity of some types of cancer,some research have demonstrated that agonist of TLR3 can trigger apoptosis of cancers.This study was proposed to investigate if Poly(I:C),the specific agonist of TLR3,could impact proliferation or apoptosis of progressive breast cancer cells MDA-MB-231,and to investigate the primary mechanism of the function.MethodsExpression of TLR1-10 mRNA was detected by quantitative real-time reverse transcription-polymerase chain reaction.Cell Counting Kit-8 was used to determine the inhibitory effect of Poly(I:C) on proliferation of MDA-MB-231 cells.Cell apoptosis was assayed by flow cytometry with V-FITC/PI staining.Results First,the toll-like receptors 1-10 were all expressed on MDA-MB-231 cells,while the expression level of TLR8 was lower than that of others.Second,according to the CCK-8,the proliferation of MDA-MB-231 cells was inhibited,but the apoptosis was not affected on the basis of Apoptosis Kit.At last,the mRNA expression of TNF-α、IFN-β and IFN-γ were elevated approximately 20 times after Poly(I:C) stimulation for 6 hours.ConclusionMDA-MB-231 cells express all toll-like receptors on mRNA level,and TLR8 was expressed lower than others.The stimulation of TLR3 with Poly(I:C) can inhibit the proliferation of MDA-MB-231,but had no effect on apoptosis.TNF-α、IFN-β and IFN-γ maybe participate in this process.
ABSTRACT
Objective Myocardial infarction and subsequent heart failure remain the most dominant health challenges worldwide.Therapeutic angiogenesis has emerged as a potential novel treatment for severe ischemic heart disease and there is increasing evidence that cell transplantation may improve the perfusion and contractility of myocardium in animal models.This study was designed to examine the endothelial growth potential and whether transplantation of human umbilical cord derived mesenchymal stem cells can improve local blood flow in a mouse ischemic hindlimb model.Methods The mesenchymal stem cells derived from human umbilical cord of passage 5 were differentiated in an endothelial differentiation medium containing vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in vitro.Samples were observed for 2 weeks.The human umbilical cord derived mesenchymal stem cells were transplanted into a hindlimb ischemia mouse model in vivo.Four weeks later,immunofluence was used to identify the migration and differentiation of the transplanted cells towards endothelial linage.Laser Doppler perfusion image was used to evaluate the local blood flow of the hindlimb.Results Results After incubation with VEGF and bFGF,the human umbilical cord derived mesenchymal stem cells started to form interconnected clusters and a network was formed.Four weeks after transplantation,the transplanted cells were sprouting f0rom the local injection and differentiated into endothelial cells,contributed to the recovery of local blood flow obviously as compared with control group.Conclusion Human umbilical cord derived mesenchymal stem cells have the ability to differentiate into endothelial cells,contribute to the local angiogenesis in a hindlimb ischemia mouse model and represent a new source for therapeutic angiogenesis for clinical applications.