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  • Cell Research
    functional studies of the relevant PI related proteins and may help shed light onto the role of PI pathways in development and cellular responses Expression of MRP14 gene is frequently down regulated in Chinese human esophageal cancer Jie WANG 1 Yan CAI 1 Hao XU 1 Jun ZHAO 2 Xin XU 1 Ya Ling HAN 1 Zhi Xiong XU 1 Bao Sheng CHEN 1 Hai Hu 1 Min WU 1 Ming Rong WANG 1 Full Text PDF Migration inhibitory factor related protein 14 MRP14 is one of calcium binding proteins referred as S100A9 The heterodimeric molecule formed by MRP14 with its partner MRP8 S100A8 is the major fatty acid carrier in neutrophils The MRP8 14 complex has been also implicated in the intracellular transport of arachidonic acid and its precursors in keratinocytes We show here the involvement of MRP14 in human esophageal cancer In an initial study mRNA differential display reverse transcription polymerase chain reaction DD PCR was performed with two esophageal carcinomas one esophageal adenocarcinoma and matched normal adjacent mucosa DD PCR with the arbitrary primer OPA3 showed that one cDNA band was highly expressed in normal tissues but disappeared or substantially decreased in tumor counterparts It was later identified to be the 3 end of migration inhibitory factor related protein 14 MRP14 Northern blotting RT PCR and Western blotting corroborated the down regulation of MRP14 in 58 64 squamous cell carcinomas and 2 2 adenocarcinomas as compared with adjacent normal epithelia of the esophagus MRP14 was undetectable in 3 3 esophageal carcinoma cell lines Immunochemistry demonstrated that expression of MRP14 was restricted to normal esophageal epithelia No mutation was found in the genomic DNA of the MRP14 gene by PCR and directed DNA sequencing Our finding suggested that the reduction of MRP14 expression is a frequent event in Chinese human esophageal cancer Impaired reproduction in transgenic mice overexpressing γ aminobutyric acid transporter I GAT1 Jia Hua HU 1 Jin Fu ZHANG 2 Ying Hua MA 1 Jie JIANG 1 Na YANG 1 Xin Bo LI 3 Zhi Guang YU CHI 4 Jian FEI 1 5 Li He GUO 1 Full Text PDF It is well documented that γ aminobutyric acid GABA system existed in reproductive organs Recent researches showed that GABA A and GABA B receptors were present in testis and sperm and might mediate the acrosome reaction induced by GABA and progesterone GABA transporter I GAT1 also existed in testis and sperm but its physiological function was unknown In the present study we used GAT1 overexpressing mice to explore GAT1 function in male reproductive system We found that the expression level of GAT1 continuously increased in wild type mouse testis from 1 month to 2 months after birth GAT1 overexpression in mouse affected testis development which embodied reduced testis mass and slowed spermatogenesis in transgenic mice Moreover transgenic mice showed increase of the percentage of broken sperm The further study revealed that the reproductive capacity was impaired in GAT1 overexpressing mice In addition testosterone level was significantly

    Original URL path: http://www.cell-research.com/artsmore.asp?id=109 (2016-02-14)
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  • Cell Research
    molecules In contrast to MTSC4 thymocyte apoptosis induced by TDCs was MHC restricted Thus MHC independent fashion of stromal DP thymocyte interaction may be one of the ways to induce thymocyte apoptosis in thymus Our study has also shown that the interaction of MTSC4 stromal cells and thymocytes is required for the induction of thymocyte apoptosis LDFF the large molecular weight DNA fragmentation factor is responsible for the large molecular weight DNA degradation during apoptosis in Xenopus egg extracts Zhi Gang LU Chuan Mao ZHANG Zhong He ZHAI Full Text PDF DNA degradation is a biochemical hallmark in apoptosis It has been demonstrated in many cell types that there are two stages of DNA fragmentation during the apoptotic execution In the early stage chromatin DNA is cut into large molecular weight DNA fragments although the responsible nuclease s has not been recognized In the late stage the chromatin DNA is cleaved further into short oligonucleosomal fragments by a well characterized nuclease in apoptosis the caspase activated DNase CAD DFF40 In this study we demonstrate that large molecular weight DNA fragmentation also occurs in Xenopus egg extracts in apoptosis We show that the large molecular weight DNA fragmentation factor LDFF is not the Xenopus CAD homolog XCAD LDFF is activated by caspase 3 The large molecular weight DNA fragmentation activity of LDFF is Mg 2 dependent and Ca 2 independent can occur in both acidic and neutral pH conditions and can tolerate 45 treatment These results indicate that LDFF in Xenopus egg extracts might be a new DNase or DNases responsible for the large DNA fragmentation ERK1 2 contributes negative regulation to STAT3 activity in HSS transfected HepG2 cells Ze Jun TIAN 2 Wei AN 1 Full Text PDF Signal transducer and activator of transcription 3 STAT3 is a recently characterized transcription factor which is essential to liver regeneration We have previously reported that hepatic stimulator substance HSS a novel growth promoting substance phosphorylated the epidermal growth factor EGF receptors and activated downstream Ras MAP kinase extracellular signal regulated kinases ERK1 2 cascade However whether HSS signal is related to STAT3 pathway remains unclear The present study is aiming to explore the regulatory effect of activation of ERK1 2 evoked by HSS on STAT3 phosphorylation and STAT3 signaling Human hepatoma cell line HepG2 was stably transfected with HSS cDNA and HSS ex pression was measured by Northern blot The results showed that the transfection of HSS into HepG2 resulted in remarkable increase in cellular proliferation as compared with the non transfected cells and it was further proved that the cellular proliferation in the HSS transfected cells was related to ERK1 2 activation Treatment of the cells with 50 μM of PD98059 an ERK1 2 specific upstream inhibitor resulted in ERK1 2 inactivation completely Inhibition of ERK1 2 allowed the tyrosine of STAT3 to be phosphorylated in a dose dependent manner to PD98059 Furthermore transient transfection of STAT3 mutant STAT3S727A into HSS bearing cells could remarkably reverse the inhibitory effect of ERK1

    Original URL path: http://www.cell-research.com/artsmore.asp?id=108 (2016-02-14)
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  • Cell Research
    were applied respectively The methylation status of the CDKN UnitName a SourceValue 2 HasSpace False Negative False NumberType 1 TCSC 0 w st on 2A promoter was determined by class SpellE methylation specific PCR and the acetylated status of the class SpellE histones associated with the p21 WAF1 and CDKN UnitName a SourceValue 2 HasSpace False Negative False NumberType 1 TCSC 0 w st on 2A genes was examined by chromatin class SpellE immunoprecipitation The expression of the CDKN UnitName a SourceValue 2 HasSpace False Negative False NumberType 1 TCSC 0 w st on 2A p21 WAF1 p53 p73 APC c myc c class SpellE Ki ras and survivin genes was detected by real time RT PCR and RT PCR The cell cycle profile was established by flow cytometry color windowtext We found that along with the demethylation of the CDKN Negative False NumberType 1 TCSC 0 w st on 2A gene promoter in both cell lines induced by 5 aza dC alone or in combination with TSA the expression of both CDKN HasSpace False Negative False NumberType 1 TCSC 0 w st on 2A and APC genes increased The treatment of TSA or sodium butyrate up regulated the transcription of p21 WAF1 significantly by inducing the acetylation of histones H4 and H3 but failed to alter the acetylation level of CDKN Negative False NumberType 1 TCSC 0 w st on 2A associated histones No changes in transcription of style mso spacerun yes p53 p73 c myc c Ki ras and survivin genes were observed In addition TSA or sodium butyrate was shown to arrest cells at the G 1 phase However 5 aza dC was not able to affect the cell cycle progression In conclusion regulation by epigenetic modification of the transcription of tumor associated genes and the cell cycle progression in both human colon cancer cell lines Colo 320 and SW1116 is gene specific Differential activation of intra S phase checkpoint in response to tripchlorolide and its effects on DNA replication Yan REN Jia Rui WU Full Text PDF DNA replication is tightly regulated during the S phase of the cell cycle and the activation of the intra S phase checkpoint due to DNA damage usually results in arrest of DNA synthesis However the molecular details about the correlation between the checkpoint and regulation of DNA replication are still unclear To investigate the connections between DNA replication and DNA damage checkpoint a DNA damage reagent tripchlorolide was applied to CHO Chinese ovary hamster cells at early or middle stages of the S phase The early S phase treatment with TC significantly delayed the progression of the S phase and caused the phosphorylation of the Chk1 checkpoint protein whereas the middle S phase treatment only slightly slowed down the progression of the S phase Furthermore the analysis of DNA replication patterns revealed that replication pattern II was greatly prolonged in the cells treated with the drug during the early S phase whereas the late replication patterns of these cells

    Original URL path: http://www.cell-research.com/artsmore.asp?id=107 (2016-02-14)
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  • Cell Research
    induced cell death while non recepter tyrosine kinase inhibitor herbimycin A HMA enhances radiation induced apoptosis In this study we focused on the modulation of radiation induced cell death by genistein and performed PCR select suppression subtractive hybridization SSH to understand its molecular mechanism We identified human thymidine kinase 1 TK1 which is cell cycle regulatory gene and confirmed expression of TK1 mRNA by Northern blot analysis Expression of TK1 mRNA and TK 1 enzymatic activity were parallel in their increase and decrease TK1 is involved in G1 S phase transition of cell cycle progression In cell cycle analysis we showed that radiation induced G2 arrest in K562 cells but it was not able to sustain However the addition of genistein to irradiated cells sustained a prolonged G2 arrest up to 120 h In addition the expression of cell cycle related proteins cyclin A and cyclin B1 provided the evidences of G1 S progression and G2 arrest and their relationship with TK1 in cells treated with radiation and genistein These results suggest that the activation of TK1 may be critical to modulate the radiation induced cell death and cell cycle progression in irradiated K562 cells Enhancement of human ACAT1 gene expression to promote the macrophage derived foam cell formation by dexamethasone Li YANG 1 Jin Bo YANG 1 Jia CHEN 1 Guang Yao YU 1 Pei ZHOU 1 Lei LEI 1 Zhen Zhen WANG 1 Catherine CY CHANG XinYing YANG 1 Ta Yuan CHANG 2 Bo Liang LI 1 Full Text PDF In macrophages the accumulation of cholesteryl esters synthesized by the activated acyl coenzyme A cholesterol acyltransferase 1 ACAT1 results in the foam cell formation a hallmark of early atherosclerotic lesions In this study with the treatment of a glucocorticoid hormone dexamethasone Dex lipid staining results clearly showed the large accumulation of lipid droplets containing cholesteryl esters in THP 1 derived macrophages exposed to lower concentration of the oxidized low density lipoprotein ox LDL More notably when treated together with specific anti ACAT inhibitors the abundant cholesteryl ester accumulation was markedly diminished in THP 1 derived macrophages confirming that ACAT is the key enzyme responsible for intracellular cholesteryl ester synthesis RT PCR and Western blot results indicated that Dex caused up regulation of human ACAT1 expression at both the mRNA and protein levels in THP 1 and THP 1 derived macrophages The luciferase activity assay demonstrated that Dex could enhance the activity of human ACAT1 gene P1 promoter a major factor leading to the ACAT1 activation in a cell specific manner Further experimental evidences showed that a glucocorticoid response element GRE located within human ACAT1 gene P1 promoter to response to the elevation of human ACAT1 gene expression by Dex could be functionally bound with glucocorticoid receptor GR proteins These data supported the hypothesis that the clinical treatment with Dex which increased the incidence of atherosclerosis may in part due to enhancing the ACAT1 expression to promote the accumulation of cholesteryl esters during the macrophage derived foam cell formation

    Original URL path: http://www.cell-research.com/artsmore.asp?id=106 (2016-02-14)
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  • Cell Research
    Whole mount in situ hybridization of TIMP 3 using DIG labeled RNA probes demonstrated that the transcripts were present in all dorsal tissues during embryogenesis but were prominent only in head structures starting at stage 35 Overexpression of TIMP 3 through transgenesis and RNA injections led to developmental abnormalities and death Both overexpression strategies resulted in post gastrulation perturbation including those to neural and head structures as well as truncated axes However RNA injections resulted in more severe early defects such as failure of neural tube closure and transgenesis caused truncated axes and head abnormalities No transgenic embryo expressing TIMP 3 survived past stage 40 The spike protein of severe acute respiratory syndrome SARS is cleaved in virus infected Vero E6 cells Xiao Dong WU 1 Bo Shang 1 Rui Fu YANG 2 Hao YU 3 Zhi Hai Ma 1 Xu SHEN 3 Yong Yong JI 1 Ying LIN 1 Ya Di Wu 1 Guo Mei Lin 1 Lin TIAN 1 Xiao Qing GAN 1 Sheng YANG 2 Wei Hong JIANG 2 Er Hei DAI 4 Xiao Yi WANG 4 Hua Liang JIANG 3 You Hua Xie 1 Xue Liang Zhu 1 Gang PEI 1 Lin LI 1 Jia Rui WU 1 Bing SUN 1 Full Text PDF Spike protein is one of the major structural proteins of severe acute respiratory syndrome coronavirus It is essential for the interaction of the virons with host cell receptors and subsequent fusion of the viral envelop with host cell membrane to allow infection Some spike proteins of coronavirus such as MHV HCoV OC43 AIBV and BcoV are proteolytically cleaved into two subunits S1 and S2 In contrast TGV FIPV and HCoV 229E are not Many studies have shown that the cleavage of spike protein seriously affects its function In order to investigate the maturation and proteolytic processing of the S protein of SARS CoV we generated S1 and S2 subunit specific antibodies Abs as well as N E and 3CL protein specific Abs Our results showed that the antibodies could efficiently and specifically bind to their corresponding proteins from E coli expressed or lysate of SARS CoV infected Vero E6 cells by Western blot analysis Furthermore the anti S1 and S2 Abs were proved to be capable of binding to SARS CoV under electron microscope observation When S2 Ab was used to perform immune precipitation with lysate of SARS CoV infected cells a cleaved S2 fragment was detected with S2 specific mAb by Western blot analysis The data demonstrated that the cleavage of S protein was observed in the lysate indicating that proteolytic processing of S protein is present in host cells Soluble expression and characterization of a GFP fused pea actin isoform PEAc1 Ai Xiao LIU Shao Bin ZHANG Xiao Jing XU Dong Tao REN Guo Qin LIU Full Text PDF A pea actin isoform PEAc1 with green fluorescent protein GFP fusion to its C terminus and His tag to its N terminus was expressed in prokaryotic cells in soluble form and

    Original URL path: http://www.cell-research.com/artsmore.asp?id=105 (2016-02-14)
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  • Cell Research
    Aβ 1 40 or Aβ 1 42 even to hundreds fold of the physiological concentration affected obviously the cell viability These results suggest that the overproduction of Aβ could not arrest cell differentiation induced by serum deprivation and that at least to a certain degree and in a limited time period is not toxic to cell viability Regulation of alternative splicing of Bcl x by IL 6 GM CSF and TPA Chang You LI 1 Jia You CHU 1 Jian Kun YU 1 Xiao Qin HUANG 1 Xiao Juan LIU 1 Li SHI 1 Yan Chun CHE 1 Jiu Yong XIE 2 Full Text PDF The splicing of many alternative exons in the precursor messenger RNA pre mRNA is regulated by extracellular factors but the underlying molecular bases remain unclear Here we report the differential regulation of Bcl x pre mRNA splicing by extracellular factors and their distinct requirements for pre mRNA elements In K562 leukemia cells treatment with interleukin 6 IL 6 or granulocyte macrophage colony stimulating factor GM CSF reduced the proportion of the Bcl xL variant mRNA while treatment with 12 O tetradecanoylphorbol 13 acetate TPA had no effect In U251 glioma cells however TPA efficiently increased the Bcl xL level These regulations were also seen for a transfected splicing reporter mini gene Further analyses of deletion mutants indicate that nucleotides 1 176 of the downstream intron are required for the IL 6 effect whereas additional nucleotides 177 284 are essential for the GM CSF effect As for the TPA effect only nucleotides 1 76 are required in the downstream intron Thus IL 6 GM CSF and TPA differentially regulate Bcl x splicing and require specific intronic pre mRNA sequences for their respective effects Dynamic tracking and mobility analysis of single GLUT4 storage vesicle in live 3T3 L1 cells Chen hong Li 1 Li Bai 1 Dong dong Li 1 Sheng Xia 1 Tao Xu 1 2 Full Text PDF Glucose transporter 4 GLUT4 is responsible for insulin stimulated glucose transporting into the insulin sensitive fat and muscle cells The dynamics of GLUT4 storage vesicles GSVs remains to be explored and it is unclear how GSVs are arranged based on their mobility We examined this issue in 3T3 L1 cells via investigating the three dimensional mobility of single GSV labeled with EGFP fused GLUT4 A thin layer of cytosol right adjacent to the plasma membrane was illuminated and successively imaged at 5 Hz under a total internal reflection fluorescence microscope with a penetration depth of 136 nm Employing single particle tracking the three dimensional subpixel displacement of single GSV was tracked at a spatial precision of 22 nm Both the mean square displacement and the diffusion coefficient were calculated for each vesicle Tracking results revealed that vesicles moved as if restricted within a cage that has a mean radius of 160 nm suggesting the presence of some intracellular tethering matrix By constructing the histogram of the diffusion coefficients of GSVs we observed a smooth distribution instead of

    Original URL path: http://www.cell-research.com/artsmore.asp?id=104 (2016-02-14)
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  • Cell Research
    molecules of TNF signaling such as RIP and TRAF2 are also involved in other cellular responses These finding suggests that RIP and TRAF2 serve a broader role than as just an effector of TNF signaling Wnt signaling in disease and in development Roel NUSSE Full Text PDF The highly conserved Wnt secreted proteins are critical mediators of cell to cell signaling during development of animals Recent biochemical and genetic analyses have led to significant insight into understanding how Wnt signals work The catalogue of Wnt signaling components has exploded We now realize that multiple extracellular cytoplasmic and nuclear components modulate Wnt signaling Moreover receptor ligand specificity and multiple feedback loops determine Wnt signaling outputs It is also clear that Wnt signals are required for adult tissue maintenance Perturbations in Wnt signaling cause human degenerative diseases as well as cancer Role of c Abl in the DNA damage stress response Yosef SHAUL Merav BEN YEHOYADA Full Text PDF c Abl has been implicated in many cellular processes including differentiation division adhesion death and stress response c Abl is a latent tyrosine kinase that becomes activated in response to numerous extra and intra cellular stimuli Here we briefly review the current knowledge about c Abl involvement in the DNA damage stress response and its implication on cell physiology Role of JNK activation in apoptosis A double edged sword Jing LIU Anning LIN Full Text PDF JNK is a key regulator of many cellular events including programmed cell death apoptosis In the absence of NF κB activation prolonged JNK activation contributes to TNF α induced apoptosis JNK is also essential for UV induced apoptosis However recent studies reveal that JNK can suppress apoptosis in IL 3 dependent hematopoietic cells via phosphorylation of the proapoptotic Bcl 2 family protein BAD Thus JNK has pro or antiapoptotic functions depending on cell type nature of the death stimulus duration of its activation and the activity of other signaling pathways Nucleo cytoplasmic communication in apoptotic response to genotoxic and inflammatory stress Jean Y J WANG Full Text PDF Genotoxic agents or inflammatory cytokines activate cellular stress responses and trigger programmed cell death We have identified a signal transduction module including three nuclear proteins that participate in the regulation of cell death induced by chemotherapeutic agents and tumor necrosis factor TNF In this nuclear signaling module retinoblastoma protein Rb functions as an inhibitor of apoptotic signal transduction Inactivation of Rb by phosphorylation or caspase dependent cleavage degradation is required for cell death to occur Rb inhibits the Abl tyrosine kinase Thus Rb inactivation is a pre requisite for Abl activation by DNA damage or TNF Activation of nuclear Abl and its downstream effector p73 induces mitochondriadependent cell death The involvement of these nuclear signal transducers in TNF induced apoptosis which does not require new gene expression indicates that nuclear events other than transcription can contribute to extrinsic apoptotic signal transduction HGF SF Met signaling in tumor progression Chong Feng GAO George F VANDE WOUDE Full Text PDF Tumor

    Original URL path: http://www.cell-research.com/artsmore.asp?id=103 (2016-02-14)
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  • Cell Research
    the epidermis In the present study we detected the expression of TGM3 in the mouse embryo using RT PCR TGM3 mRNA is weakly presented from E11 5 to E14 5 and increases significantly from E15 5 to birth Then we determined the spatial and temporal expression pattern of TGM3 in the skin and other organs by in situ hybridization We found a deprivation of TGM3 in skin at E11 5 while a rich supply in periderm cells and a weak expression in basal cells from E12 5 to E14 5 From the period of E15 5 to E16 5 after keratinization in the epidermis TGM3 was expressed in the granular and cornified layers The electron microscopic observation of the C57BL 6J mouse limb bud skin development provided several morphological evidences for the epidermal differentiation The above findings suggest that the expression of TGM3 plays a important role in the epidermis differentiation in embryogenesis Down regulation of IL 8 expression in human airway epithelial cells through helper dependent adenoviral mediated RNA interference Huibi CAO 1 4 Anan WANG 1 Bernard MARTIN 1 David R KOEHLER 1 4 Pamela L ZEITLIN 5 A Keith TANAWELL 1 2 3 Jim HU 1 2 4 Full Text PDF Interleukin IL 8 is a potent neutrophil chemotactic factor and a crucial mediator in neutrophil dependent inflammation Various cell types produce IL 8 either in response to external stimuli such as cytokines or bacterial infection or after malignant transformation Anti IL 8 strategies have been considered for anti inflammatory therapy In this paper we demonstrate that the RNA interference technique can be used to efficiently down regulate IL 8 protein expression in airway epithelial cells We used a helper dependent adenoviral vector to express a small hairpin sh RNA targeting human IL 8 in cultured airway epithelial cells IB3 1 Cftr C38 Cftr corrected stimulated with TNF α IL 1β or heat inactivated Burkholderia cenocepacia Stimulated IL 8 expression in IB3 1 and C38 cells was significantly reduced by shRNA expression The shRNA targeting IL 8 had no effect on the activation of NF κB or on the protein levels of IkB or IL 6 suggesting that this anti IL 8 strategy was highly specific and therefore may offer potential for the treatment of inflammatory diseases Dynamic distribution of Ser 10 phosphorylated histone H3 in cytoplasm of MCF 7 and CHO cells during mitosis Deng Wen LI Qin YANG Jia Tong CHEN Hao ZHOU Ru Ming LIU Xi Tai HUANG Full Text PDF The dynamic distribution of phosphorylated Histone H3 on Ser10 phospho H3 in cells was investigated to determine its function during mitosis Human breast adenocarcinoma cells MCF 7 and Chinese hamster cells CHO were analyzed by indirect immunofluorescence staining with an antibody against phospho H3 We found that the phosphorylation begins at early prophase and spreads throughout the chromosomes at late prophase At metaphase most of the phospho H3 aggregates at the end of the condensed entity of chromosomes at equatorial plate During anaphase

    Original URL path: http://www.cell-research.com/artsmore.asp?id=102 (2016-02-14)
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