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MAWBP and MAWD inhibit proliferation and invasion in gastric cancer

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World Journal of Gastroenterology 2013年第18期19卷 2781-2792页

作者： Dong-Mei Li Jun Zhang Wen-Mei Li Jian-Tao Cui Yuan-Ming Pan Si-Qi Liu Rui Xing You-Yong Lu Laboratory of Molecular Oncology Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education) Peking University Cancer Hospital/Institute Department of Biochemistry and Molecular Biology School of Medicine Shihezi University Beijing Institute of Genomics Chinese Academy of Sciences

AIM: To investigate role of putative mitogen-activated protein kinase activator with WD40 repeats (MAWD)/ MAWD binding protein (MAWBP) in gastric cancer (GC). METHODS: MAWBP and MAWD mRNA expression level was examined by real-time reverse transcriptasepolymerase chain reaction and semi-quantitative polymerase chain reaction in six GC cell lines. Western blotting was used to examine the protein expression levels. We developed GC cells that stably overexpressed MAWBP and MAWD, and downregulated expression by RNA interference assay. Proliferation and migration of these GC cells were analyzed by 3-(4,5-dimethyl2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT), soft agar, tumorigenicity, migration and transwell assays. The effect of expression of MAWBP and MAWD on transforming growth factor (TGF)-β1-induced epithelialmesenchymal transition (EMT) was examined by transfection of MAWBP and MAWD into GC cells. We detected the levels of EMT markers E-cadherin, N-cadherin and Snail in GC cells overexpressing MAWBP and MAWD by Western blotting. The effect of MAWBP and MAWD on TGF-β signal was detected by analysis of phosphorylation level and nuclear translocation of Smad3 using Western blotting and immunofluorescence. RESULTS: Among the GC cell lines, expression of endogenous MAWBP and MAWD was lowest in SGC7901 cells and highest in BGC823 cells. MAWBP and MAWD were stably overexpressed in SGC7901 cells and knocked down in BGC823 cells. MAWBP and MAWD inhibited GC cell proliferation in vitro and in vivo . MTT assay showed that overexpression of MAWBP and MAWD suppressed growth of SGC7901 cells (P < 0.001), while knockdown of these genes promoted growth of BGC823 cells (P < 0.001). Soft agar colony formation experiments showed that overexpression of MAWBP and MAWD alone or together reduced colony formation compared with vector group in SGC7901 (86.25±8.43, 12.75±4.49, 30±6.41 vs 336.75±22.55, P < 0.001), and knocked-down MAWBP and MAWD demonstrated opposite effects (131.25±16.

关键词： Gastric cancer Mitogen-activated protein kinase activator with WD40 repeats binding protein Mitogen-activated protein kinase activator with WD40 repeats Invasion Transforming growth factor-β Epithelial-mesenchymal transition

Hepatic regeneration and the epithelial to mesenchymal transition

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World Journal of Gastroenterology 2013年第9期19卷 1380-1386页

作者： Zeng-Fu Xue Xiu-Min Wu Ming Liu Department of Digestive Diseases The First Affiliated Hospital of Xiamen University Xiamen 361003 Fujian Province China Department of Pharmacy The First Affiliated Hospital of Xiamen University Xiamen 361003 Fujian Province China

Liver injuries are repaired by fibrosis and regeneration. The core stage is the repair response and fibrosis formation as a scar. The cause of overly-responsive scar formation and diminished regeneration, especially in liver fibrosis and cirrhosis, is still unknown. The epithelial to mesenchymal transition (EMT), a previously discovered mechanism, plays an important role in liver fibrosis and tumor metastasis. Recently, EMT has been found to be associated with liver and bile duct cell fibrosis. Analyzing the established models and chronic disease processes, we propose that EMT liver cells may also lose their regenerative capability due to phenotype changes and that the remaining liver cells may quickly lose their regenerative capability in liver fibrosis or cirrhosis. Recognizing these phenotype changes or transition cells may play an important role in targeting therapy to reverse fibrosis not only by disrupting the transition that is necessary to produce the extracellular matrix but also by restoring the regenerative capacity of EMT-like cells.

关键词： Epithelial to mesenchymal transition Hepatocyte Regeneration Fibrosis Transforming growth factor-β Liver Epithelial to mesenchymal transition -like Hepatocyte stellate cells

Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice

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World Journal of Gastroenterology 2013年第32期19卷 5250-5260页

作者： Kazuhiro Takahashi Soichiro Murata Kiyoshi Fukunaga Nobuhiro Ohkohchi Department of Surgery University of Tsukuba

AIM:To investigate the role of human platelets in liver ***:Severe combined immunodeficiency(SCID)mice were administered CCl4and either phosphate-buffered saline(PBS group)or human platelet transfusions(hPLT group).Concentrations of hepatocyte growth factor(HGF),matrix metallopeptidases(MMP)-9,and transforming growth factor-β(TGF-β)in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay(ELISA)and Western *** effects of a human platelet transfusion on liver fibrosis included the fibrotic area,hydroxyproline content,and-smooth muscle actin(α-SMA)expression,which were evaluated by picrosirius red staining,ELISA,and immunohistochemical staining using an anti-mouse-SMA antibody,*** of mesenchymal-epithelial transition factor(Met)and SMAD3,downstream signals of HGF and TGF-β,were compared between the two groups by Western blotting and were quantified using *** apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end ***,the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 ***:The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group(fibrotic area,1.7%±0.6%vs 2.5%±0.6%,P=0.03;hydroxyproline content,121±26 ng/g liver vs 156±47 ng/g liver,P=0.04).There was less α-smooth muscle actin staining in the hPLT group than in the PBS group(0.5%±0.1%vs 0.8%±0.3%,P=0.02).Hepatic expression levels of mouse HGF and MMP-9were significantly higher in the hPLT group than in the PBS group(HGF,109±13 ng/g liver vs 88±22 ng/g liver,P=0.03;MMP-9,113%±7%/GAPDH vs 92%±11%/GAPDH,P=0.04).In contrast,the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group(22±5ng/g

关键词： Human platelet Liver fibrosis Hepatocyte apoptosis Hepatocyte growth factor Transforming growth factor-β Matrix metallopeptidases

Role of LAP^+CD4^+ T cells in the tumor microenvironment of colorectal cancer

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World Journal of Gastroenterology 2017年第3期23卷 455-463页

作者： Wu Zhong Zhi-Yuan Jiang Lei Zhang Jia-Hao Huang Shi-Jun Wang Cun Liao Bin Cai Li-Sheng Chen Sen Zhang Yun Guo Yun-Fei Cao Feng Gao Department of Gastrointestinal Surgery Ganzhou Hospital of Nanchang University Department of Anorectal Surgery Minzu Hospital of Guangxi Zhuang Autonomous Region Department of Colorectal and Anal Surgery First Affiliated Hospital of Guangxi Medical University

AIM To investigate the abundance and potential functions of LAP^+CD4^+ T cells in colorectal cancer(CRC). METHODS Proportions of LAP^+CD4^+ T cells were examined in peripheral blood and tumor/paratumor tissues of CRC patients and healthy controls using flow cytometry. Expression of phenotypic markers such as forkhead box(Fox)p3, cytotoxic T-lymphocyte-associated protein(CTLA)-4, chemokine CC receptor (CCR)4 and CCR5 was measured using flow cytometry. LAP^-CD4^+ and LAP^+CD4^+ T cells were isolated using a magnetic cellsorting system and cell purity was analyzed by flow cytometry. Real-time quantitative polymerase chain reaction was used to measure expression of cytokines interleukin (IL)-10 and transforming growth factor(TGF)-β.RESULTS The proportion of LAP^+CD4^+ T cells was significantly higher in peripheral blood from patients (9.44% ± 3.18%) than healthy controls (1.49% ± 1.00%, P < 0.001). Among patients, the proportion of LAP^+CD4^+ T cells was significantly higher in tumor tissues(11.76% ± 3.74%) compared with paratumor tissues (3.87% ± 1.64%, P < 0.001). We also observed positive correlations between the proportion of LAP^+CD4^+ T cells and TNM stage(P < 0.001), distant metastasis(P < 0.001) and serum level of carcinoembryonic antigen(P < 0.05). Magnetic-activated cell sorting gave an overall enrichment of LAP^+CD4^+ T cells (95.02% ± 2.87%), which was similar for LAP^-CD4^+ T cells(94.75% ± 2.76%). In contrast to LAP^-CD4^+ T cells, LAP^+CD4^+ T cells showed lower Foxp3 expression but significantly higher levels of CTLA-4, CCR4 and CCR5(P < 0.01). LAP^+CD4^+ T cells expressed significantly larger amounts of IL-10 and TGF-β but lower levels of IL-2, IL-4, IL-17 and interferon-γ, compared with LAPCD4+ T *** LAP^+CD4^+ T cells accumulated in the tumor microenvironment of CRC patients and were involved in immune evasion mediated by IL-10 and TGF-β.

关键词： LAP^+CD4^+ T cells Colorectal cancer Tumor microenvironment Interleukin-10 Transforming growth factor-β

Toxicarioside A,isolated from tropical Antiaris toxicaria,blocks endoglin/TGF-βsignaling in a bone marrow stromal cell line

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Asian Pacific Journal of Tropical Medicine 2012年第2期5卷 91-97页

作者： Yue-Nan Li Feng-Ying Huang Wen-Li Mei Hao-Fu Dai Jun-Li Guo Guang-Hong Tan Peng Zhou Agriculture College Hainan University Hainan Provincial Key Laboratory of Tropical Medicine Hainan Medical College Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences

Objective:To investigate possible mechanism of toxicarioside A in HS-5 bone stromal ***:HS-5 bone stromal cells were cultured in media supplemented with various concentrations of toxicarioside A or control DMSO(not treatment).Endoglin and TGF-βwere detected by Northern and Western blot analysis and quantified in a standard method. Downstream molecules of endoglin and TGF-β(Smad1,Smad2 and their active phosphorylated counterparts,pSmad1 and pSmad2) were also detected and quantified by Western blot analysis. In addition,cell proliferation assay and small interfering RNA(siRNA) against endoglin were used to certificate the function of endolgin in the HS-5 ***:Compared with the not treated(0μg/mL) or DMSO treated control HS-5 cells,HS-5 cells treated with toxicarioside A were found significant attenuation of endolgin and TGF-β*** inhibition of cell proliferation was also found in the HS-5 cells treated with toxicarioside ***1-related Smad1 and ALK5-related Smad2 were decreased in HS-5 cells treated with toxicarioside *** addition,phosphorylated Smad1(pSmad1) and Smad2(pSmad2) were also found attenuation in toxicarioside A-treated HS-5 *** interference showed that blockage of endoglin by siRNA also decreased Smad1 and Smad2 expression in HS-5 ***:Our results indicate that toxicarioside A can influence bone marrow stromal HS-5’s function and inhibit HS-5 cell proliferation by alteration of endoglin-related ALK1(Smad1) and ALK5(Smad2) signaling.

关键词： Toxicarioside A Endoglin Transforming growth factor-β Signal pathway Cell proliferation

BMP-4 induced proliferation and oriented differentiation of rat hepatic oval cells into hepatocytes

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Asian Pacific Journal of Tropical Medicine 2015年第5期8卷 412-416页

作者： Zhi-Ming Wang Xiao-Hua Yuan Hong Shen Department of Hepatobiliary Surgery Xiangya HospitalCentral South University Institute of Medical Science Xiangya HospitalCentral South University

Objective:To explore the role of bone morphogenetic protein 4(BMP-4) in hepatic progenitor cells(HPCs).Methods:The effect of BMP-4 on rat hepatic oval cells was examined by using the WB-F344 rat hepatocytic epithelial stem-cell-like cell *** hepatocytic cell line could exert various hepatocytc functions including the secretion of albumin and *** was used to examine the effects of BMP-4 and its antagonist,Noggin,on the proliferation and differentiation of these cells,cellular uptake and excretion of indocyanine green,the periodic acid-schiff(PAS) assay for glycogen storage and the expression of hepatic ***:Our results showed for the first time that BMP-4 may acted as a potential inducer of hepatic differentiation in rat hepatic oval ***:This cell source offers a much-needed attractive and expandable source for future investigations of drug screening,stem cell technologies and cellular transplantation,in a society with increasing levels of liver disease and damage.

关键词： Bone morphogenetic protein-4 Transforming growth factor-β Hepatic progenitor cells Proliferation Differentiation

Treg/Th17 cell balance and phytohaemagglutinin activation of T lymphocytes in peripheral blood of systemic sclerosis patients

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World Journal of Experimental Medicine 2017年第3期7卷 84-96页

作者： Ekaterina Krasimirova Tsvetelina Velikova Ekaterina Ivanova-Todorova Kalina Tumangelova-Yuzeir Desislava Kalinova Vladimira Boyadzhieva Nikolay Stoilov Tsvetelina Yoneva Rasho Rashkov Dobroslav Kyurkchiev Laboratory of Clinical Immunology University Hospital "St. Ivan Rilski"Department of Clinical Laboratory and Clinical ImmunologyMedical University of Sofia Clinic of Rheumatology University Hospital "St. Ivan Rilski"Department of Internal MedicineMedical University of Sofia

AIM To investigate T-cell activation, the percentage of peripheral T regulatory cells(Tregs), Th17 cells and the circulating cytokine profile in systemic sclerosis(SSc).METHODS We enrolled a total of 24 SSc patients and 16 healthy controls in the study and divided the patients as having diffuse cutaneous SSc(dc SSc, n = 13) or limited cutaneous SSc(lc SSc, n = 11). We performed a further subdivision of the patients regarding the stage of the disease-early, intermediate or late. Peripheral venous blood samples were collected from all subjects. We performed flow cytometric analysis of the activationcapacity of T-lymphocytes upon stimulation with PHA-M and of the percentage of peripheral Tregs and Th17 cells in both patients and healthy controls. We used ELISA to quantitate serum levels of human interleukin(IL)-6, IL-10, tissue growth factor-β1(TGF-β1), and IL-17 *** We identified a decreased percentage of CD3+CD69+ cells in PHA-stimulated samples from SSc patients in comparison with healthy controls(13.35% ± 2.90% vs 37.03% ± 2.33%, P < 0.001). However, we did not establish a correlation between the down-regulated CD3+CD69+ cells and the clinical subset, nor regarding the stage of the disease. The activated CD4+CD25+ peripheral lymphocytes were represented in decreased percentage in patients when compared to controls(6.30% ± 0.68% vs 9.36% ± 1.08%, P = 0.016). Regarding the forms of the disease, dc SSc patients demonstrated lower frequency of CD4+CD25+ T cells against healthy subjects(5.95% ± 0.89% vs 9.36% ± 1.08%, P = 0.025). With regard to Th17 cells, our patients demonstrated increased percentage in comparison with controls(18.13% ± 1.55% vs 13.73% ± 1.21%, P = 0.031). We detected up-regulated Th17 cells within the lc SSc subset against controls(20.46% ± 2.41% vs 13.73% ± 1.21%, P = 0.025), nevertheless no difference was found between dc SSc and lc SSc patients. Flow cytometric analysis revealed an increased percentage of CD4+CD25-Foxp3+ in dc SSc patients c

关键词： Systemic sclerosis T-cell activation Th17 Tregs CD4+CD25-Foxp3+cells Interleukin-17 Tissue growth factor-β Interleukin-10 Interleukin-6

Role of IL-10 in the progression of kidney disease

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World Journal of Transplantation 2013年第4期3卷 91-98页

作者： Inna Sinuani Ilia Beberashvili Zhan Averbukh Judith Sandbank Pathology Department Assaf Harofeh Medical CenterSackler Faculty of MedicineTel Aviv UniversityZerifin70300Israel Nephrology Division Assaf Harofeh Medical CenterSackler Faculty of MedicineTel Aviv UniversityZerifin70300Israel

Interleukin-10(IL-10), a cytokine with anti-inflammatory and immunomodulatory functions, regulates the biology of B and T cells. The present review describes the role of IL-10 in normal renal physiology, during acute kidney injury and in the development of chronic renal failure. We further discuss IL-10-induced cellular and molecular pathways and their link to the progression of kidney injury.

关键词： Transforming growth factor-β Cystatin C Interleukin-10 receptor End-stage renal disease Mesangial cell proliferation Epithelial-to-mesenchymal transition Allograft rejection

Genetic expression of Col-2A and Col-10A as a function of administration of IGF-1 &TGF-<i>β</i>with and without anterior mandibular repositioning appliance on the growth of mandibular condylar cartilage in young rabbit

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Open Journal of Stomatology 2013年第9期3卷 6-13页

作者： A. S. Patil R. B. Sable R. M. Kothari P. Nagarajan Deartment of Orthodontics and Dentofacial Orthopedics Bharati Vidyapeeth Dental College and Hospital Bharati Vidyapeeth Deemed University Pune India Rajiv Gandhi Institute of Biotechnology Bharati Vidyapeeth Deemed University Pune India

New Zealand (NZ) young rabbits with the administration of insulin-like growth factor (IGF-1) and transforming growth factor-β (TGF-β) with and without mandibular anterior repositioning appliances are explored for the growth of the mandibular condylar cartilage (MCC). 32 growing NZ and rabbits were divided into 4 groups: the group with saline injection in TMJ, the group which received growth factor injection in TMJ, the group which received anterior positioning appliance and the group which received growth factors injection as well as mandibular repositioning appliance. Gene expression was studied by real-time RT-PCR and cartilage growth by histomorphometry. Administration of growth factors along with mandibular repositioning appliances has induced 1) 1.70-fold expression of Col-2Agene (p value factors only induced 1) mere 0.56-fold expression of Col-2Agene (p value 10A gene (p value growth factors along with mandibular repositioning appliances causes an increase in genetic expressions which have been corroborated by histomorphometry and validated by statistical analysis, during an accelerated growth of mandibular condylar cartilage. Administration of growth factors in the TMJ could provide a synergistic role along with mandibular repositioning appliances for treatment of mandibular retrognathism as well as disorders on the MCC.

关键词： Transforming Growth factor-β (TGF-β) Insulin-Like Growth factor (IGF-1) Condylar Cartilage Growth Mandibular Repositioning Appliances Col-2A Col-10A