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Induction of CXC chemokines in human mesenchymal stem cells by stimulation with secreted frizzled-related proteins through non-canonical Wnt signaling

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World Journal of Stem Cells 2015年第11期7卷 1262-1273页

作者： David S Bischoff Jian-Hua Zhu Nalini S Makhijani Dean T Yamaguchi Research Service VA Greater Los Angeles Healthcare System Department of Medicine David Geffen School of Medicine at UCLA

AIM: To investigate the effect of secreted frizzledrelated proteins(s FRPs) on CXC chemokine expression in human mesenchymal stem cells(h MSCs).METHODS: CXC chemokines such as CXCL5 and CXCL8 are induced in h MSCs during differentiation with osteogenic differentiation medium(OGM) and may be involved in angiogenic stimulation during bone repair. h MSCs were treated with conditioned medium(CM) from L-cells expressing non-canonical Wnt5 a protein, or with control CM from wild type L-cells, or directly with s FRPs for up to 10 d in culture. m RNA expression levels of both CXCL5 and CXCL8 were quantitated by real-time reverse transcriptase-polymerase chain reaction and secreted protein levels of these proteins determined by ELISA. Dose-(0-500 ng/m L) and time-response curves were generated for treatment with s FRP1. Signal transduction pathways were explored by western blot analysis with pan- or phosphorylation-specific antibodies, through use of specific pathway inhibitors, and through use of si RNAs targeting specific frizzled receptors(Fzd)-2 and 5 or thereceptor tyrosine kinase-like orphan receptor-2(Ro R2) prior to treatment with s FRPs. RESULTS: CM from L-cells expressing Wnt5 a, a noncanonical Wnt, stimulated an increase in CXCL5 m RNA expression and protein secretion in comparison to control L-cell CM. s FRP1, which should inhibit both canonical and non-canonical Wnt signaling, surprisingly enhanced the expression of CXCL5 at 7 and 10 d. Dickkopf1, an inhibitor of canonical Wnt signaling prevented the s FRPstimulated induction of CXCL5 and actually inhibited basal levels of CXCL5 expression at 7 but not at 10 d post treatment. In addition, all four s FRPs isoforms induced CXCL8 expression in a dose- and time-dependent manner with maximum expression at 7 d with treatment at 150 ng/m L. The largest increases in CXCL5 expression were seen from stimulation with s FRP1 or s FRP2. Analysis of mitogen-activated protein kinase signaling pathways in the presence of OGM sh

关键词： CXC chemokines Mesenchymal stem cell Osteogenesis differentiation Wnt signaling pathway Frizzled-related protein Frizzled receptors

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Effects of Hexamethylene Bisacetamide on Proliferation and Gene Expression of Human Hepatocarcinoma Cells

Effects of Hexamethylene Bisacetamide on Proliferation and G...

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第十三届中国科协年会第6分会场-绿色经济与沿海城市可持续发展战略研讨会

作者： Lanying CHEN 1,Haibo YANG 1,Jie SUN 1 ,Zhaohui XIE 1,Qifu LI 2(1. Henan University of Urban Construction, department of Biology Engineering Pingdingshan 467044, China 2.The Key Laboratory of China Education Ministry for Cell Biology and Tumor Cell Engineering,School of Life Sciences,Xiamen University,Xiamen 361005, China)

(AIM)To investigate the effects of 5 mmol/L hexamethylene bisacetamide (HMBA) on proliferation and gene expression of human hepatocarcinoma SMMC-7721 cells.(METHODS)Effects of HMBA on proliferation of SMMC-7721 cells were determined with trypan blue dye exclusion test and HE staining. Effects of HMBA on cell cycle were detected by flow cytometry. Effects of HMBA on cell morphology by Light microscope and electron microscope. Protein levels of c-fos, c-myc, bcl-2, c-erbB-2, p53, rb, p21 and p27 were examined by immunocytochemistry.(RESULTS)The proliferation of SMMC-7721 cells were inhibited after 5 mmol/L HMBA treatment, and the inhibitory rate of cellular growth is 52.4 %. HMBA treatment could markedly increase the proportion of cells at G0/G1 phase and decrease the proportion at S phase. light microscope and electron microscope showed that SMMC-7721 cells had undergone the restorational alteration in morphology and ultrastructure after HMBA treatment. The expression levels of oncogene c-fos, c-erbB-2, c-myc,bcl-2 and mtp53 proteins were down-regulated while the expression level of tumor suppressor gene rb, p21 and p27 proteins were up-regulated after the treatment with 5 mmol/L HMBA. (CONCLUSIO)NHMBA could effectively inhibit the proliferation of hepatocarcinoma cells, reverse the malignant morphological and ultrastructural characteristics, alter the levels of enzymes and antigens, regulate the expression of differentiation-associated oncogene and tumor suppressorgene, and induce hepatocarcinama cell differentiation.

关键词： biology HMBA human hepatocarcinoma cell differentiation

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Dissection of Developmental Mechanisms of Neurocristopathy in Waardenburg Syndrome and Piebaldism Mouse Models

Dissection of Developmental Mechanisms of Neurocristopathy i...

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第二届全国发育生物学大会

作者： Ling Hou Developmental Cell Biology and Disease Program Vision Science Research CenterWenzhou Medical University

Many human neurocristopathies include genetic defects in a number of syndromes,tumors,and dysmorphologies of neural crest stem cell derivatives during *** coat color mutations have long served as an excellent model for the study of neural crest stem cell developmental *** to shared embryonic origin of various tissues or to pleiotropic effects,these mutants also serve as human disease models for many aspects of studies including vision,hearing,craniofacial abnormalities,digestive disorders,and neural tube *** recent years a complex network of genes has been associated with pigmentation abnormalities and deafness in neurocristopathies resulting from NC stem cell-derived melanocyte *** example,Waardenburg syndrome(WS)can result from mutations either in SoxlO,Mitf or Ednrb and Piebaldism can result from mutations in *** studies have revealed surprising new insights into how this network of interacting genes plays complex functional roles in melanocyte ***,we are using Waardenburg Syndrome and Piebaldism mouse models to dissect developmental mechanisms of *** this point of view,I will provide some of our findings and explore potential mechanisms by which this regulatory network of interacting genes may control melanocyte precursor differentiation.

关键词： genetic disease neural crest stem cell gene interaction melanocyte differentiation

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Astrocytes cultured on water-repellent fractal tripalmitin surface demonstrate superior morphology and functions compared with traditional culture substrate

Astrocytes cultured on water-repellent fractal tripalmitin s...

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中国神经科学学会第十届全国学术会议

作者： Zhe WANG Wei-Wei HU Bi-Yuan HONG Shan-Shan ZHANG Wen-Yun FANG Zhong CHEN Department of Pharmacology Key Laboratory of Medical Neurobiology of the Ministry of Health of China Zhejiang Province Key Laboratory of Neurobiology College of Pharmaceutical Sciences Department of Chemistry Zhejiang University

Objective In the CNS,astrocytes play an essential role with their multiple functions and sophisticated structure,as surrounded by a fractal environment which hasn’t been available in our traditional cell *** sought to investigate whether a water-repellent fractal nanomaterial surface made by tripalmitin(PPP),could provide the cultured primary astrocytes with an improved closer-to-natural environment for experimental assessment of glial structure and *** Tripalmitin is a hydrophobic triglyceride derived from palmitic acid with good biocompatibility and can form water-repellent fractal structure during the phase transition from the metastable α-form to the stable β-crystalline *** examined morphology and biochemical characterization of astrocytes cultured 3 days on poly-1-lysine(control) and PPP surfaces with contact angles of 105o15°(non-fractal but water-repellent),125°35°(fractal and water-repellent) and 150°160°(fractal and super water-repellent).Results We found that(1)water-repellent fractal PPP surface can induce astrocytes to display differentiated morphology with smaller size of cell area,longer and finer filopodium-like processes,and higher morphological complexity revealed by fractal *** super water-repellent fractal PPP surface with water contact angle of 150°60° produced the maximal effects compared with other surfaces at lower water contact angle.(2) The trends of characteristic protein expression,including that of nestin,vimentin,GFAP and glutamine synthetase,for astrocytes cultured in super water-repellent fractal PPP surface approximated more to in vivo pattern compared with those in conventional cell culture surface.(3) This super water-repellent PPP surface also rendered astrocytes to perform more pronounced promotion of neurogenesis by increasing the release of nerve growth factor in a co-culture *** Our results suggest that super water-repellency fractal PPP surface facilitates the astrocytes to mimi

关键词： astrocyte differentiation fractal tripalmitin water-repellency

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Wnt5a induced bone destruction through activating JNK in pre-osteoblasts and MSCs of collagen-induced arthritis arthritic mice

Wnt5a induced bone destruction through activating JNK in pre...

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第十一届全国免疫学学术大会

作者： Niu menglin Tong Yulong Du Yuxuan Yuan Huihui Zhao Wenming Department of Immunology School of Basic Medical SciencesCapital Medical University Department of Blood Transfusion Peking University Cancer Hospital & Institute

Background:This study aimed to detect the expression of Wnt5 a in the hind paws bone joint tissue of collagen-induced arthritis(CIA) mice and to evaluate the effects and mechanisms of Wnt5 a on pre-osteoblastic cells(MC3T3-E1 cells) and bone mesenchymal stem cells(MSCs) in secretion and *** and Results:In vivo,Wnt5 a was overexpression in the hind paw joints upon CIA in *** immunofluorescent staining showed that the high level expression of Wnt5 a was mainly localized in osteoblasts from the CIA mice compared to normal controls,which relating to the increasing osteoblasts *** vitro,compared with Wnt3 a treated group,adding Wnt5 a reduced the expression of OPG and increased the IL-6 level in MC3T3-E1 cells and MSCs,while no significantly changes were determined in the expression of IL-1β,TNF-α and *** addition,Wnt5 a treatment induced the MC3T3-E1 cells activation and expression of alkaline phosphatase(ALP) and Runt-related transcription factor 2(RUNX2) in both mR NA and ***,the Ror2 mR NA expression was also higher in Wnt5 a treated cells than ***,Wnt5 a up-regulated JNK signal protein expression and decreased significantly β-*** results demonstrated that the up-regulating of Wnt5 a in osteoblasts caused synovitis,bone erosion and abnormal new bone formation through changing the levels of OPG,IL-6,ALP and ***,Wnt5 a possible bound its receptor Ror2,stimulated the JNK signal pathway and suppressed the canonical Wnt pathway to aggravate arthritis in CIA ***:The study suggested that Wnt5 a closely contributed to the pathogenesis of rheumatoid arthritis(RA).

关键词： Wnt5a CIA Pre-osteoblast Secretion differentiation RA

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Establishment of a novel culture method for adult myoblasts of rat amplification in batches

Establishment of a novel culture method for adult myoblasts ...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Shu-Hong LIU~(1 *) Yong-Qi ZHAO~1 Hai-Tao WU~1 Xiao-Ping CHEN~2 Ming FAN~1 1 Institute of Basic Medical Sciences Academy of military Medical Sciences Beijing 10085 2 China Astronaut Research and Training Center

＜正＞Objective To establish a method to rapidly isolate myoblasts from adults and amplify in *** Myoblasts were isolated from adult skeletal muscle by using "One Step Digestion with Mixture Enzyme" method we previously founded;then,purified and amplified in batches with novel method so called "Purification with Diverse Adherence Method".Results In the new isolation method,the cell isolation time was 2.10.52 h,which was markedly reduced compared to 6.80.67 h for the old isolation method（P＜0.05）.The myf5/desmin positive rate in isolated myoblasts was more than 97%purified with "Purification with Diverse Adherence Method".The characteristics of the isolated myobalts by the new method had a preferable proliferation potency and were not different from that by the old method in regard to differentiate into myotubes in differentiation media（P＞0.05）.Conclusion It could be used to rapidly isolate and obtain highly purified myoblasts by the novel established isolation and amplification in batches methods.

关键词： myoblasts amplification in batches proliferation differentiation myotubes

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Tetramethylpyrazine as a potential promotion for neurogenesis in neural stem cell involved in mitogen-activated protein kinases in hypoxia disease

Tetramethylpyrazine as a potential promotion for neurogenesi...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yu-Mei TIAN,Yong LIU~*,Xin-Lin CHEN,Hai-Xia ZHANG,Jun-Feng ZHANG,Qin-Dong SHI,Peng-Bo YANG Institute of Neurobiology,Environment and Genes Related to Diseases Key Laboratory of Education Ministry,the State Key Subject for Physiology,Xi''an Jiaotong University College of Medicine

＜正＞Objective This study investigated the effects of tetramethylpyrazine（TMP）,an active element of tradional Chinese medicine Ligusticum Chuanxiong,on proliferation and differentiation of neural stem cells（NSCs） from rat brain in hypoxia condition and the activation of mitogen-activated protein kinases（MAPKs） signaling pathway during the processes. Methods we use the rat NSCs culture to investigate the effects of TMP on cell proliferation and differentiation with MTT and immunocytochemistry and western blotting analysis,and the the phosphorylation of ERK,JNK and p38 in MAPK signaling pathway was examined after TMP *** The results showed that TMP increased the proliferation and promoted the differentiation of the NSCs into *** increased the phosphorylation of ERK1/2 and decreased the phosphorylation of p38 at different *** inhibitor（U0126） in part blocked the defferetiation of the NSCs into neurons induced by *** Our findings demonstrated that TMP enhanced the proliferation and differentiation of NSCs of rat after hypoxia in vitro,in which the alteration of phosphorylation of ERK and p38 is involved.

关键词： tetramethylpyrazine neural stem cell proliferation differentiation mitogen-activated protein kinases

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Differential effects of Kriippel-like transcriptional factors on the regulation of endothelial progenitor cells

Differential effects of Kriippel-like transcriptional factor...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Xiao-Xia LI~1,Yi-Meng SONG~1,Cheng-Lai FU~1,Zhen-Jiu ZHU~1,Ying-Ying HAN~1,Cheng-Hong LI~1,Nan-Ping WANG~(2,*),Yi ZHU~(1,*) 1 Department of Physiology and Pathophysiology 2 Institute of Cardiovascular Research,Key Laboratory of Cardiovascular Science of Ministry of Education,Peking University Health Science Center,Beijing 100191,China

Objective:Endothelial progenitor cells(EPCs) participate in post-natal *** previously reported that AMPK activation promoted EPC ***,the transcriptional regulation of EPC differentiation and proliferation remains largely *** this study,we investigated the potential roles of Kr(u|¨)ppel-like transcription factors(KLFs) in the functions of *** and Results:We found that following the prolong culture of human cord blood-derived late EPCs,the expression of KLF2 was *** of AMPK by VEGF,AMPK agonist and lovastatin promotes EPC differentiation,and KLF2 upregulation was much more obvious. On the other hand,co-treatment of a high dose of VEGF and leukemia inhibitory factor(LIF) activated Akt and increased EPCs proliferation,the level of KLF2 being decreased,but KLF4 being increased *** effects of VEGF/LIF could be inhibited by DN-AKT adenovirus ***,overexpression of KLF4 with adenovirus enhanced the expression of CD34,but not increased the phosphorylation of Akt and mTOR,suggesting Akt activation is the upstream of KLF4 *** of KLF2 with RNAi in EPCs decreased the ability of tube formation of EPCs in vitro,indicating the important role of KLF2 in EPC-mediated ***,we found that bone marrow-derived EPCs from eNOS mice had strong growth ability and more CD34 positive cells, compared with those from wild-type mice,but showed weak tube formation *** blot results revealed a higher level of KLF4 and lower level of KLF2 in eNOS ***:Our results demonstrate an opposite effect of two members of KLFs on the proliferation and differentiation of EPCs,which may provide new insights into the regulation of EPC-mediated angiogenesis.

关键词： endothelial progenitor cells proliferation differentiation eNOS AMPK Akt bone marrow

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Bone Marrow Stromal Cells Differentiate into Mature Neural Cells in vitro.

Bone Marrow Stromal Cells Differentiate into Mature Neural C...

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2011中华医学会神经外科学学术会议

作者： Yiwu DAI1 1.Bayi Brain Hospital, Beijing Military General Hospital

To Explore the feasibility of differentiation into neural cells from bone marrow stromal cells (BMSCs) in vitro, and to offer reference for the applying of BMSCs in the field of neuroscience. Methods BMSCs were acquir... 详细信息

关键词： Adult bone marrow mesenchymal stem cells (ABMMSC) differentiation Neuronal cells Glial cells.

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Development of cancer-initiating cells and immortalized cells with genomic instability

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World Journal of Stem Cells 2015年第2期7卷 483-489页

作者： Ken-ichi Yoshioka Yuko Atsumi Hitoshi Nakagama Hirobumi Teraoka Division of Carcinogenesis and Cancer Prevention National Cancer Center Research Institute Department of Pathological Biochemistry Medical Research InstituteTokyo Medical and Dental University

Cancers that develop after middle age usually exhibit genomic instability and multiple mutations. This is in direct contrast to pediatric tumors that usually develop as a result of specific chromosomal translocations andepigenetic aberrations. The development of genomic instability is associated with mutations that contribute to cellular immortalization and transformation. Cancer occurs when cancer-initiating cells(CICs), also called cancer stem cells, develop as a result of these mutations. In this paper, we explore how CICs develop as a result of genomic instability, including looking at which cancer suppression mechanisms are abrogated. A recent in vitro study revealed the existence of a CIC induction pathway in differentiating stem cells. Under aberrant differentiation conditions, cells become senescent and develop genomic instabilities that lead to the development of CICs. The resulting CICs contain a mutation in the alternative reading frame of CDKN2A(ARF)/p53 module, i.e., in either ARF or p53. We summarize recently established knowledge of CIC development and cellular immortality, explore the role of the ARF/p53 module in protecting cells from transformation, and describe a risk factor for genomic destabilization that increases during the process of normal cell growth and differentiation and is associated with the downregulation of histone H2 AX to levels representative of growth arrest in normal cells.

关键词： ARF/p53 module Cancer stem cells Cancerinitiatingcells differentiation Genomic instability H2AX

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