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Objective To investigate the relation between vasculogenic mimicry (VM) and MIG-7 in osteosarcoma, as well as their roles in the prognosis, and to establish a model for predicting the prognosis of osteosarcoma. Methods VM was identified by CD31/PAS double-staining in 156 cases of AJCC stage Ⅱ extremity osteosarcoma. Tumor samples were also immunohistochemically stained for MIG-7 to determine whether it was associated with the occurrence of VM. Univariate and multivariate Cox regression analyses were used to identify prognostic factors and a prognostic nomogram for predicting 3- and 5-year OS and MFS was constructed. C-index and calibration curves were used to verify the predictive accuracy of the model. Results The MIG-7 expression in osteosarcoma tissues was associated with VM formation, but MIG-7 expression was not associated with gender, age, AJCCⅡA/ⅡB stage, tumor location, surgical type or histological response to pre-operative chemotherapy. Survival analysis showed that MIG-7 expression, VM and pre-operative chemotherapy were identified as three independent prognostic factors. The value of C-index in nomogram was greater than 0.7. The predicted calibration curve was similar to the standard curve. Conclusion MIG-7 accelerates the progression of osteosarcoma by promoting VM formation, and may also affect prognosis through other mechanisms. The nomogram could afford accurate prognosis prediction and individualized diagnosis and treatment for osteosarcoma patients.
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Introduction@#More than 30 years have passed since the revived and developed of traditional medicine, and dozens of textbooks and publications on traditional medicine have been published and qualified personnel have been trained. However, there is a need to drastically improve the quality of traditional medical care and the knowledge and skills of doctors. Therefore, it is important to translate, study books written by ancient physicians, maaramba, and scholars, and to apply theory, diagnosis, and treatment methods in training and practice.@*Research materials and methods@#Basic research materials: Naiman toin Jambaldorj (1792-1855). གསོ་བྱེད་མཛས་མཚར་ མིག་རྒྱན། Tibetan scripture of a wooden printing block.@*Method@#We used germenevtic method and checklist method.@*Result@#In the frame of this research, we used the medical sutra written by Jambaldorj as the main material in order to clarify the specialty and number of animal-oriented medicines in the “mdzes mtshar mig rgyan”. During the research, it was possible to clarify the structure, general content, and specialty of the sutra written by Jambaldorj, classify animal-oriented medicines according to the cyrillic alphabet, and make allegory names for some of the raw materials of animal origin. In addition, the study of animal-oriented medicines in the sutra showed that many issues were important, such as their identification, usages, and new production. As a result of the first study, we identified the characteristics and usages of some essential animal-oriented medicines.@*Conclusion@#Jambaldorj began writing the “mdzes mtshar mig rgyan” before leaving for Tibet, and later, during his pilgrimage to Tibet, he enriched and completed it in his book. It was found to correspond to the period 1817-1823. A selection of 124 animal-oriented medicines from the book “mdzes mtshar mig rgyan” was made. In this way, it is possible to interpret and explain the hidden meanings of animal-oriented medicines in this sutra, and to determine the usages of pharmacology and drug formulations.
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OBJECTIVE@#To investigate the inhibitory effects of silencing migration-inducing gene-7 (Mig-7) on vasculogenic mimicry formation, migration and invasion of human glioma cells and whether MEK/ERK signaling pathway mediates these effects.@*METHODS@#Human glioma U251 cells were infected by lentiviral vectors carrying a small interfering RNA targeting Mig-7 gene (sh-Mig-) or a negative control shRNA (sh-NC), and real-time quantitative PCR was used to detect the expression level of Mig- mRNA in the cells. Three-dimensional culture and Transwell chamber invasion assay were used to observe the effect of Mig- gene silencing on vasculogenic mimicry formation and invasion ability of the U251 cells. Western blotting was performed to detect the changes in the protein expression levels of MEK/ERK in the infected cells.@*RESULTS@#We successfully obtained a U251 cell line with stable low expression of Mig- gene using RNA interference technique. Compared with the cells infected with sh-NC lentivirus and the non- infected cells, U251 cells infected with the lentiviral vector carrying sh-Mig- showed significantly decreased expression level of Mig- ( < 0.01) with obviously lowered vasculogenic mimicry formation and invasion abilities ( < 0.05). Mig- silencing also significantly lowered the expressions of MEK and ERK proteins in U251 cells ( < 0.05).@*CONCLUSIONS@#Silencing of Mig-7 gene inhibits vasculogenic mimicry formation and invasion of U251 cells possibly by suppressing MEK/ERK signaling, suggesting the important role of Mig-7 gene in vasculogenic mimicry formation and invasion of human glioma cells.
Subject(s)
Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Silencing , Glioma , Genetics , Pathology , Neoplasm Proteins , Metabolism , RNA, Small Interfering , Signal TransductionABSTRACT
Mig1 and Snf1 are two key regulatory factors involved in glucose repression of Saccharomyces cerevisiae. To enhance simultaneous utilization of glucose and xylose by engineered S. cerevisiae, single and double deletion strains of MIG1 and SNF1 were constructed. Combining shake flask fermentations and transcriptome analysis by RNA-Seq, the mechanism of Mig1 and Snf1 hierarchically regulating differentially expressed genes that might affect simultaneous utilization of glucose and xylose were elucidated. MIG1 deletion did not show any significant effect on co-utilization of mixed sugars. SNF1 deletion facilitated xylose consumption in mixed sugars as well as co-utilization of glucose and xylose, which might be due to that the SNF1 deletion resulted in the de-repression of some genes under nitrogen catabolite repression, thereby favorable to the utilization of nitrogen nutrient. Further deletion of MIG1 gene in the SNF1 deletion strain resulted in the de-repression of more genes under nitrogen catabolite repression and up-regulation of genes involved in carbon central metabolism. Compared with wild type strain, the MIG1 and SNF1 double deletion strain could co-utilize glucose and xylose, and accelerate ethanol accumulation, although this strain consumed glucose faster and xylose slower. Taken together, the MIG1 and SNF1 deletions resulted in up-regulation of genes under nitrogen catabolite repression, which could be beneficial to simultaneous utilization of glucose and xylose. Mig1 and Snf1 might be involved in the hierarchical regulatory network of genes under nitrogen catabolite repression. Dissection of this regulatory network could provide further insights to new targets for improving co-utilization of glucose and xylose.
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Chemokine CXCL9/Mig (monokine induced by IFN-?) belongs to the subfamily of chemotactic cytokines known as CXC-chemokines. In vivo CXCL9 is mainly induced by IFN-? in macrophages and primary glial cells. In vitro, CXCL9 can be secreted by cells such as macrophages, microvascular endothelial cells and neutrophils, in response to the synergy of IFN-? and TLR(toll-like receptor) ligands. CXCL9 is a chemoattractant for activated T lymphocytes, tumor-infiltrating T-lymphocytes, but not for neutrophils or monocytes. The receptor specific for CXCL9 is CXCR3, a G protein-coupled protein which has seven transmembrane domain. The structure and the chemical characterization of CXCL9, as well as its effects on autoimmune deseases, allograft rejection, cancer therapy were reviewed.
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BACKGROUND: The pathogenesis of acute myeloid leukemia (AML) is complicated by DNA damage, balanced or unbalanced translocation, deletion, inversion, abnormal transcription factors, receptors, and others. The CCAAT/enhancer binding protein alpha (C/EBPalpha) and C/EBP epsilon (C/EBPepsilon), one of transcription factors, play important roles in normal granulopoiesis. We wished to assess whether increasing the activity of either C/EBPalpha or C/EBPepsilon could suppress the leukemic myeloblasts. METHODS: To make retrovirus, BOSC23 cells were transfected with retroviral constructs; mouse stem cell retrovirus-internal ribosomal entry site-green fluorescent protein (MIG), MIG-C/EBP alpha-estrogen receptor (ER) and MIG-C/EBP epsilon-ER. 32Dcl3 murine myeloblastic (32Dcl3) cells or #1111 acute promyelocytic leukemic (#1111 APL, #1111) cells were transduced with each retrovirus. Growth rate and differential cell count were examined, and granulocytic surface markers of Gr-1 and Mac-1 were checked. Transduced #1111 cells were injected into 20 sublethally irradiated (4.5 Gy) mice; at day 14, 4 groups of 5 mice each were input into subcutaneous tissue with placebo, 4-hydroxytamoxifen (4HT), all trans retinoic acid (ATRA), or 4HT & ATRA pellets; survival times were analysed when they died. RESULTS: The number of GFP (+) transduced 32Dcl3 cells with MIG (control group) at days 2, 4, and 6 were 684976, 1975965, and 2808244; 32Dcl3 cells with MIG-C/EBP alpha-ER were 77354, 53180, and 39460; and 32Dcl3 cell with MIG-C/EBP epsilon-ER were 328384, 698424, and 974850, respectively. The control group didn't express both Gr-1 and Mac-1, but C/EBP alpha expressed 56.1%, 55.6% and C/EBPepsilon expressed 31.3% and 32.6%, respectively. The differential counts of immature, intermediate, and mature forms in control group were 90.0%, 6.0%, and 4.0%; C/EBP 4.3%, 33.7%, and 62.0%; C/EBPepsilon 41.0%, 48.3%, and 10.7%, respectively. The mean survival time of transduced #1111 cells with MIG-C/EBP alpha-ER injected mice was 30.5 days in placebo group, 41.8 days in 4HT (C/EBP ) group, 69.0 days in ATRA group, and 97.8 days in 4HT (C/EBP ) & ATRA group. In case of MIG-C/EBP epsilon-ER, the survival time was 26.4 days in placebo group, 33.0 days in 4HT (C/EBP ) group, 49.6 days in ATRA group, and 52.5 days in 4HT (C/EBP ) & ATRA group. CONCLUSIONS: Both C/EBP and C/EBP suppressed cell growth and differentiation of 32Dcl3 cells, and they also suppressed cell growth of #1111 cells. The ATRA was more effective than C/EBP in APL, and C/EBP and ATRA had synergistic effects in APL. The growth arrest and differentiated action of C/EBPalpha was more powerful than that of C/EBPepsilon.
Subject(s)
Animals , Mice , Carrier Proteins , Cell Count , DNA Damage , Granulocyte Precursor Cells , Leukemia, Myeloid, Acute , Retroviridae , Stem Cells , Subcutaneous Tissue , Survival Rate , Transcription Factors , Tretinoin , ZidovudineABSTRACT
Interferon-gamma (IFN-gamma) is well known as a potent inducer in monokine induced by IFN-gamma (Mig) mRNA expression. Although lipopolysaccharide (LPS) alone is weakly effective on Mig mRNA expression. the stimulation of LPS and IFN-gamma (LPS/IFN-gamma simultaneously has been shown to synergize to produce a high level of Mig mRNA in mouse peritoneal macrophages. In this study, interleukin-10 (IL-10) was found to suppress the LPS/IFN-gamma- induced Mig mRNA expression in cell type- and mouse strain-specific fashion, but IFN-gamma alone-induced Mig mRNA was unaffected by IL-10 under identical experimental conditions. The IL-10-mediated suppression of LPS/IFN-gamma-stimulated Mig mRNA expression was dependent on the concentration of IL-10, and was prevented when the agent was added 2 hours after LPS/IFN-gamma treatment. The suppressive action of IL-10 was dependent on a protein synthesis. However, IL-10 did not reduce the stability of LPS/IFN-gamma-induced Mig mRNA. These data may have important implications for a previously unrecognized role for IL-10 as a regulator of synergistic effect of LPS on the IFN-gamma-induced expression of the Mig gene in macrophages.
Subject(s)
Animals , Mice , Gene Expression , Interferon-gamma , Interleukin-10 , Macrophages , Macrophages, Peritoneal , RNA, MessengerABSTRACT
BACKGROUND: Kawasaki disease is an acute febrile illness with systemic vasculitis which primarily affects children, We examined the production of leptin in plasma and gene expressions of CXC chemokines in peripheral blood mononuclear cells from patients with Kawasaki disease. METHODS: Consecutive 39 samples from 13 patients according to the different clinical stages (acute, subacute, convalescent) of Kawasaki disease were collected. The plasma leptin levels according to clinical stages of Kawasaki disease were examined by ELISA and the expression of IP-10, Mig and IL-8 mRNAs in 39 samples (13 samples of each stage) from 13 cases were examined by RT-PCR. RESULTS: There were not significant changes of plasma leptin levels according to the clinical stages of Kawasaki disease. The mean values of plasma leptin concentrations during each of the stages (n=13, p>0.05, pg/ml) were 335.8+/-549.0 in acute, 358+/- 347.6 in subacute, and 443.6+/-645.9 in convalescent stage. The mRNAs of IP-10, Mig, and IL-8 were expressed in 13/13 (100%), 2/13 (15%), 9/13 (69%) during acute stage, 13/13 (100%), 6/13 (46%), 13/13 (100%) during subacute stage, and 13/13 (100%), 4/13 (31%), 10/13 (77%) during the convalescent stage, respectively. In three patients, the production of leptin and expression of IP-10 mRNA were dramatically decreased according to the process of the clinical stages. In five patients with prominent cervical lymphadenopathy, the expression of IL-8 mRNA during the subacute stage was more elevated than the acute and convalescent stages. CONCLUSION: This data suggests that the production of leptin and the gene expressions of IP-10, Mig and IL-8 seem to have no significant correlation to the clinical stages of Kawasaki disease. However, expression patterns of IP-10, Mig and IL-8 mRNA may be related to the specific clinical manifestations, and the expression of IP-10 may also be correlated to leptin levels with pericardial involvement.
Subject(s)
Child , Humans , Chemokines, CXC , Enzyme-Linked Immunosorbent Assay , Gene Expression , Interferon-gamma , Interleukin-8 , Leptin , Lymphatic Diseases , Mucocutaneous Lymph Node Syndrome , Plasma , RNA, Messenger , Systemic VasculitisABSTRACT
Lipoarabinomannans (LAM) is believed as a potential virulence factor of Mycobacterium tuberculosis. LAM exhibits marked differences in biological activities depending on the types, arabinofuranosyl-terminated LAM (AraLAM) derived from a rapidly growing Mycobacterium sp. and heavily mannosylated LAM (ManLAM) derived from the Erdman strain. Collaboration between macrophages and T cells, especially macrophage activation by gamma interferon (IFN-r) and chemoattraction of T cells at the very inflammatory foci would be essential in defence against M. tubercu/osis. Chemokines Mig and IP-10 are inducible by IFN-r from macrophages and have been shown to act in vitro as T cell chemoattractants. However, little is known of LAMs capacity to induce chemokines Mig and IP-10 in macrophages. In this experiment, Mig and IP10 mRNA was expressed in the delayed-type hypersensitivity (DTH) against BCG in BCG-immune mice. In some experiments, both Mig and IP-10 mRNA was evidently induced with different time courses in THP-1 cells stimulated with whole live M. tubercu/osis H37Rv (Erdman). To investigate whether Mig and IP-10 genes are differentially induced depending on the type of LAM, PCR amplification was used to detect mRNA of Mig and IP-10 from the THP-1 human monocytic cells stimulated with LAM. AraLAM, but not ManLAM, induced weakly Mig and IP-10 mRNA in the THP-1 cells. The induction of Mig and IP-10 was dependent upon the dose of AraLAM and exhibited different time courses. The mRNA for Mig and IP-10 was induced within 2 hr and 4 hr from the initiation of treatrnent and has disappeared by 8 hr and 24 hr under the experimental conditions used in this study, respectively. IFN-y at 100 U/ml, but not at 10 U/ml, was itself a good stimulus of both Mig and IP- 10 expression, and synergized with either AraLAM or ManLAM for induction of both Mig and IP-10. The expression patterns of MCP-3 were somewhat similar to those of Mig and IP10 in all of the experiments. These data indicate that IFN-r may contribute to effective macrophage function if macrophages are not fully affected by ManLAM, and chemokines Mig and IP-10 may a role in recruitment of T cells at inflammatory foci of tuberculosis.
Subject(s)
Animals , Humans , Mice , Chemokines , Chemotactic Factors , Cooperative Behavior , Hypersensitivity , Interferons , Macrophage Activation , Macrophages , Mycobacterium , Mycobacterium bovis , Mycobacterium tuberculosis , Polymerase Chain Reaction , RNA, Messenger , T-Lymphocytes , Tuberculosis , VirulenceABSTRACT
"Mig is a gamma interferon-inducible T cell chemoattractant that is a member of the chemokine family of cytokines. In order to gain a better understanding of the molecular mechanisms that regulate expression of the Mig gene, we have characterized the Mig gene and compared its structure and regulatory sequences with that of its ciosest IP10 gene. The genomic organization of the Mig gene reveals three introns that interrupt the transcribed sequence into four functional domains with a single ""CAT""- and ""TATA""-like structure. Primer extension analysis was used to identify the transcriptional initiation site that is located 50 bp upstream to the methionine codon that begins the long open reading frame. Comparison of the intron-exon structure of this gene to the gene for IP10 establishes that both genes are interrupted in precisely the same positions within homologous codons. The similarity of the intron-exon structure of the Mig and IP10 genes further support the hypothesis that Mig and IP10 genes have evolved from a common ancestral gene by gene duplication. The 5'-flanking region of Mig gene shows no overall sequence similarity with that from its closest IP10 gene whose production is also affected by gamma interferon. However, there are regions including a sequence with similarity to the NFxB binding site, AP-1 binding site, and ISRE. The r-RF-1 binding site is well conserved from -204 to -194 from the transcription start site in the Mig gene. Given the importance of IFN-r for effective immunity in tuberculosis and induction of Mig and IP10 genes in macrophages by IFN-r, we demonstrated induction of the genes Mig and IP10 with different message levels in the THP-1 human monocytic cell lines stimulated with whole M. tuberculosis. Despite the very similarity in genomic organization and the overlap in biological activities between MIG and IP10, our data described herein further support the suggestion that these chemokines rnay role nonredundantly in vivo. Moreover, our studies done on the Mig gene should provide the structural framework for future studies and begin to dissect cis-acting DNA sequences that are critical for gene regulation mediated by cell surface receptors."
Subject(s)
Humans , Base Sequence , Binding Sites , Cell Line , Chemokine CXCL9 , Chemokines , Codon , Cytokines , Gene Duplication , Genome , Interferons , Introns , Macrophages , Methionine , Open Reading Frames , Transcription Factor AP-1 , Transcription Initiation Site , TuberculosisABSTRACT
Objective:To study the inhibitory effect of IL-27 against human tumor angiogenesis and the related mechanisms.Methods: Human esophageal carcinoma cells(Eca109/IL-27) stably transfected with IL-27 gene were injected into nude mice to establish tumor-bearing mouse model.The survival time and tumor growth were observed.IFN-? level secreted by splenocytes was measured by ELISA.Expression of VEGF and CD34 was detected by immunohistochemistry method and MVD was calculated according to CD34 level.RT-PCR was used to detect the expression of IP-10 and MIG mRNA in the tumor tissues.Results: The survival time of mice injected with Eca109/IL-27 cells was significantly longer than those of mice injected with wide type Eca109 or Eca109/LXSN(blank vector) cells(P