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Ribophorin II is a substrate of spermatid-specific E3 ligase RNF148

Ribophorin II is a substrate of spermatid-specific E3 ligase...

引用

“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Wei Zhang~(1,2),Zhenyu Xiao~(1,2),Chunsheng Han~1 1.State Key Laboratory of Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101,China 2.Graduate University of Chinese Academy of Sciences,Beijing 100039,China

Ring finger proteins(RNFs) are a subfamily of zinc finger proteins and many of them function as ubiquitin ligases(E3s).Many RNFs are germ cell-specifically expressed and play important roles in spermatogenesis. Previously,we have shown that RNF133 is a testis-specific endoreticulum-associated *** this study,we show that another spermatid-specific gene Rnf148,located 5k-bp upstream of Rnfl33,also encodes an E3 with similar protein domains including a signal peptide,a protease-association domain,a transmembrane domain and a C3HC4 RING finger *** vitro assays showed that RNF148 worked with ubiquitin-conjugating enzymes(E2s) such as UbcH5b and UbcH7,which is involved in endoreticulum associated degradation(ERAD) ***148 co-localized with the reticulum-associated E2 MmUbc6, and co-immunoprecipitated with CD3-.which is a typical substrate in ERAD *** two-hybrid screening revealed that RNF148 physically interacts with ribophorin II(RPNII),which is a component of ***-immunoprecipitation and cycloheximide treatment experiments confirmed that RPNII is not only a partner but also a substrate of *** was highly expressed in spermatogonia and spermatocytes,in which RNF148 was *** negative correlation of the expression of RNF148 and RPNII in mouse testis implied that RPNII is targeted by RNF148 for degradation in *** RPNII is involved in the biosynthesis process of N-glycans on glycoproteins, which has been reported to play key roles in germ cell-Sertoli cell attachment,we propose that RNF148 may inhibit the synthesis of glycoproteins through the degradation of RPNII resulting in the dissociation of germ cell from Sertoli cells and their release into the seminiferous tubules.

关键词： RNF148 RPNII RING E3 ligase ubiquitination spermatogenesis

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Expression of an Intrabody against Human Cyclin Dl Provides Antitumor Efficacy on Hepatocellular Carcinoma both in vitro and in vivo

Expression of an Intrabody against Human Cyclin Dl Provides ...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Bingzhi Li,Dan Yang,Hongliang Xu,Wenhan Wang,Yan Wu,Guiying Li Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education,Jilin University,Changchun,130021,China

Cyclin D1 is a rate-limiting key factor in controlling the transition from Gl to S in cell *** and overexpression of Cyclin D1 occurs in tumorigenesis of several types of human cancer including hepatocellular carcinoma(HCC).It becomes a potential target for the treatment of ***,by linking appropriate intracellular trafficking signal sequences to their coding genes,can be directed to all intracellular compartments to neutralize and block the function of target *** study was designed to use an intrabody against cyclin D1 to inhibit the growth and proliferation of HCC cells through blocking and inhibiting the biological activity of cyclin D1 overexpressed in cancer *** expression plasmid pER-ADK habouring an endoplasmic reticulum(ER)-retained scFv gene against human Cyclin D1(ER-ADκ) was *** pER-ADK was transfected into HepG2 *** RT-PCR and western blotting showed the anti-cyclinD1 intrabody was expressed efficiently in HepG2 *** assay showed that ER-ADk was localized in the endoplasmic *** results suggested that ER-ADk could recognize and combine its target protein cyclinD1 of HepG2 *** assay showed that ER-ADk significantly inhibited the growth of HepG2 cells(P<0.01).The results of FACS showed that anti-cyclin D1 intrabody arrested most of HepG2 cells at G1 phase and distinctly induced apoptosis of HepG2 cells(P<0.01).ER-ADk expression also suppressed the growth of subcutaneous tumors in nude mice in vivo after the intratumoral injection of *** study suggested that anti-cyclin D1 intrabody inhibits the growth and proliferation of HCC cells both in vitro and in vivo by blocking cyclin D1 activity and it might be a potential approach for hepatocellular carcinoma therapy.

关键词： cyclin D1 intrabody HCC gene therapy

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Early Growth Response-1 Accelerates Liver Regeneration after Partial Hepatectomy through GGPPS in Mice

Early Growth Response-1 Accelerates Liver Regeneration after...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Lai Shanshan~1,Shen Ning~2,Li Chaojun~2,Xue Bin~2 1.The Medical School of Nanjing University,Nanjing 210093, 2.China Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing Normal University,Nanjing 210097,China

Background & Aims:Liver regeneration comprises a series of complicated processes,in which several transcription factors play an important ***-1 is rapidly induced as part of the immediate-early gene response of the regenerating *** studies indicate that EGR-1-/- exhibits delayed hepatocellular mitotic progression after partial hepatectomy and increased liver injury after carbon tetrachloride(CC14) *** the mechanism of Egr-1 in regulates regeneration progress is still *** aim is to reveal an important signal pathway mediated by Egr-1 in this ***:70%partial hepatectomy(PH) were performed using mice tail vein injection of adenoviral vectors expressing GFP (Ad-GFP),Egr-l(Ad-Egr-l) or Dominant negative Egr-l(Ad-DNEgr-l) to increase or down-regulate hepatic Egr-1 levels,and mice lack of ggpps in hepatocyte(GGPPS-/-).Animals were sacrificed at intervals after the surgery,and the remnant liver was harvested and *** in vitro study,serum starvation recovery model was used to mimic liver regeneration process in normal human *** signal pathway was detected by western ***:Loss of Egr-1 function in mice liver caused a delayed liver regeneration process reflected by regeneration ratio and expression of cell-cycle related proteins, increased ALT and AST level and ***-expressed Egr-1 can promoted GGPPS transcription increased Ras prenylation and finally caused Erk1/2 activation in ***-1/GGPPS/ ERK pathway contributes to the expression of cyclins in serum or partial hepatectomy induced cell cycle of hepatocytes. GGPPS-/- mice also shows decreased PCNA,CyclinD and CyclinE ***:Egr-1 plays an important role in regulating the cell cycle of hepatocytes induced by partial ***-1 contributes to the expression of cyclins through activating ggpps transcription,which leads to the activation of Ras/Raf/MAPK ***-1 gives positive feedback to MAPK pathway,whi

关键词： early growth response 1 liver regeneration GGPPS

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Functionally mature blood cells derived from human pluripotent stem cells:Possible rule in future clinical utilization.

Functionally mature blood cells derived from human pluripote...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Feng MA Institute of Blood Transfusion,Chinese Academy of Medical Sciences(CAMS) & Peking Union Medical College 26 Huacai Road,DongSanHuanErDuan,Chengdu 610052,China

The establishment of human embryonic stem cell(hESC) and induced pluripotent stem cells (hiPSC) has greatly expanded our knowledge about the early development in human *** the past decade,hESCs and hiPSCs have been proved excellent tools in characterization of molecular and cellular mechanisms controlling the normal and diseased differentiation of hematopoietic progenitors and mature, functional blood *** with the advances in research,a clinical translation of hESC/hiPSC-derived hematopoietic cells as novel therapies has been foreseen in near *** our laboratory,we recently established efficient blood cell-inducing systems by co-culture of hESC/hiPSCs with murine fetal stromal cells derived from aorta-gonad-mesonephros(AGM) region and fetal *** the co-culture, undifferentiated hESC/hiPSC colonies grow up and differentiate to firstly a mesoderm like structure then hematopoietic progenitor cells on days 10 to *** the second culture system,these hESC/hiPSC-derived hematopoietic progenitors are further induced along to some specific blood cell lineages,such as erythrocytes,mast cells,and eosinophils,*** a clone-tracing method,we gained concrete results that hESC-derived erythrocytes kept continuously progressing toward maturation over a time *** expression of P-globin vs e-globin showed a typical switching pattern,mimicking the normal development of human *** day 12 of the co-culture with murine fetal liver-derived stromal cells, hESC-derived erythrocytes expressβ-globin at about 60%,but upregulated to almost 100%with additional 6 days of *** mature hESC-erythrocytes can undergo enucleation and release oxygen(PNAS 105:13087;2008).The unlimited source and the robust growth of hESC/hiPSC-derived erythrocytes hint a brilliant possibility as the substitute for transfusion *** the other hand,hESC/hiPSC-derived erythrocytes can be ideal models for uncovering the mechanisms controlling human erythrocyte diseas

关键词： Huamn ES and iPS cells Hematopoiesis Differentiation Erythrocytes Mast Cells Eosinophiles

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A preliminary investigation on the role of miR-185 in breast cancer

A preliminary investigation on the role of miR-185 in breast...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Rui Wang~1,Hong-Fei Xia~2 and Xu Ma~2 1.Graduate School of Peking Union Medical College,Beijing,100005 2.Reproductive and Genetic Center of National Research Institute for Family Planning,Beijing,100081

MicroRNA are endogenously small non-coding RNAs which regulate gene expression by blocking the translation or decreasing the stability of ***-185 is one of this family members,which can induce cell cycle arrest in human non small cell lung cancer and suppress tumor ***'s still unclear whether it has the relationship to breast cancer or *** this study,we found that miR-185 inhibited cellular proliferation and promoted apoptosis in T47D cells,indicating that miR-185 might inhibit the occurrence of breast *** futher explore the possible mechanisms in this process,we analysed its target genes. Targetscan and PicTar algorithms showed that stathmin 1(STMN1) may be one of miR-185 target genes. STMN1 is a microtubule-associated protein involved in the regulation of the microtubule filament system by destabilizing microtubules and takes part in the preventative progression of ER-positive primary in breast *** validate STMN1 as the target of miR-185,we set up a luciferase reporter assay.3'-UTR segment of STMN1 was amplified by PCR from human cDNA and inserted downstream of the termination codon of the luciferase reporter gene in the pGL3 control vector.3'-UTR segment of STMN1,deleting putative miR-185 biding site,was used as a negative *** sense reporter vector was respectively co-transfected with miR-185 mimics,mimics negative control,miR-185 inhibitors,or inhibitor negative control into T47D *** results shows that luciferase activity was not affected neither miR-185 mimics nor miR-185 inhibitors,which indicates that STMN1 may not be the target gene of *** further identify this results,the sense or control reporter vector was co-transfected with miR-185 ***, the luciferase activity wasn't changed *** results indicate that miR-185 may have no association with ***,our data collectively suggest that miR-185 may be involved in the preventative progression of bresast cancer,not related with STMN1.

关键词： miR-185 breast cancer STMN1 target genes

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Expression and function of Zfp261 in spermatogenesis

Expression and function of Zfp261 in spermatogenesis

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Shangying Liao,Yujian Wu,Chunsheng Han State Key Laboratory of Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing,China 100101

The proliferation and differentiation of spermatogonial stem cells(SSCs) are the driving force for *** elucidation of gene regulations and interactions are essential to understand this complex *** cell-derived neurotropic factor(GDNF) has been recognized as an indispensible growth factor for the proliferation of *** mining of GDNF-regulated genes,we have identified Zfp261 to be down-regulated as consistently reported by three different ***261 encodes a putative protein of 1370 amino acids,containing ten putative zinc finger *** protein sequences are highly conserved among *** has been reported that disruption of Zfp261 by chromosome translocation could be the cause for X-linked mental retardation(XLMR).Zfp261 is ubiquitously expressed in embryonic and adult tissues with the highest levels in the brain and the ***261 is present in a transcriptional repressive complex that also contains ZNF198,LSD1,CoREST and *** on these data,we hypothesize that Zfp261 plays an important role in mammalian *** the present study,we demonstrated that ZFP261 was detected in both germ cells and somatic cells in mouse *** it was predominantly present in nuclear of undifferentiated spermatogonia and sex body in spermatocytes,it was also weakly expressed in Sertoli *** using lentivirus-mediated RNA inference (RNAi),we showed that the knockdown of Zfp261 could promote the proliferation of ***,the effect of RNAi is independent of GDNF,implying that Zfp261 could be a main downstream mediator of the GDNF signaling *** a first step to explore the molecular mechanism of ZFP261 function,we used the yeast two-hybrid screening,co-immunoprecipitation(Co-IP),immunoprecipitation-mass spectrometry(IP/MS) techniques to identify its interactive *** putative partners of ZFP261 that we have discovered include several transcription factors such as TATA-Binding Protein 1(TBP1), TBP-

关键词： SSCs Zfp261 immunohistochemistry RNAi Yeast two-hybrid system immunoprecipitation mass spectrometry

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Identification and tissue expression of four interleukin-7 mRNA transcripts in Chinese tree shrew(Tupaia belangeri)

Identification and tissue expression of four interleukin-7 m...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Xiao-Hong Liu~(1,2),Yu Fan~(1,2),Yong-Gang Yao~1 1.Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences & Yunnan Province,Kunming Institute of Zoology,32 Jiaochang Donglu,Kunming,Yunnan 650223,China 2.Graduate School of the Chinese Academy of Sciences,Beijing 100039,China

Interleukin-7(IL7) is a pleiotropic cytokine mat is actively involved in immune *** plays an important role in the development,survival and expansion of T cells,B cells,Dendritic cells and natural killer *** tree shrew,a small experimental animal with a close affinity to primate,has been proposed as an alternative experimental animal to primates in biomedical ***,there is a lack of basic knowledge about the immune system of tree *** this study,we characterized the expression pattern of IL7 gene in different tissues of Chinese tree *** used reverse transcription polymerase chain reaction(RT-PCR) and rapid amplification of cDNA end(RACE) approaches to clone the entire IL7 gene in tree *** then quantified the expression profile of IL7 mRNA transcripts in eight tissues (including heart,liver,spleen,lung,kidney,intestine,skeletal muscle and brain) in 20 tree shrews using quantitative real-time PCR.A total of 6 mRNA transcripts were discerned from the cDNA samples from different *** canonical form of IL7 gene is 1817 bp in length and encodes 177 amino acids. Besides this transcript,we found that three transcripts,termed as IL7-svl,IL7-sv2,IL7-sv3,were highly expressed in all tissues except for *** IL7-sv3 had significantly high expression in skeletal *** relatively high expression level of IL7 mRNA in different tissues is in accordance with the immuno-regulation function of ***,the exact role of different IL7 mRNA transcripts remains to be determined.

关键词： interleukin-7 tree shrew mRNA transcript tissue expression

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Association and molecular mechanism between apoptotic regulation and chemosensitivity of esophageal cancer

Association and molecular mechanism between apoptotic regula...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Yang Xu~1,Lanping Zhou~1,Jing Huang~1,Fang Liu~1,Jian Yu~2,Qimin Zhan~1,Lin Zhang~2,Xiaohang Zhao~1 1.State Key Laboratory of Molecular Oncology,Cancer Institute & Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100021,China 2.University of Pittsburgh Cancer Institute,and Departments of Pathology and Pharmacology and Chemical Biology,University of Pittsburgh School of Medicine,Pittsburgh,PA 15213,USA

Object:To examine the expression of Smac,a proapoptotic gene,and c-IAP1,an apoptotic inhibitor protein,in the esophageal squamous cell carcinoma tissues,and investigate the molecular mechanisms of Smac and c-IAP1 regulaiton of ***:In current study,immunohistochemistry and western blot methods were used to detect the expression of Smac and c-IAP1 in tissues from ESCC and adjacent *** and gene transfection were used to select a stable clone with Smac knockdwon. Cell cytotoxity response to CDDP and long-term survival were evaluated via MTT assay, immuno-fluoresence and colony *** rate and cellular substructure detected by transmission electron microscope(TEM) and flow-cytometry with Annexin V-PI staining,apoptosis related proteins and molecular mechanism withwestern blot *** mimetics on chemosensitivity of ESCC were *** therapeutic response of ESCC cells with different Smac status in vivo were compared using xenograft ***:We found that Smac was significantly down-regulated in esophageal cancer(64.7%,44/68,P=0.001),and Smac expression was significantly different(P<0.05) between chemosensitive(with CR or PR responses) and chemoresistant(with SD or PD) *** associations of tested factors with response was tested using logistic regression *** ESCC cells treated with cisplatin,a commonly used chemotherapeutic drug,Smac and cytochrome C were released from mitochondria,in company with caspase-3 and caspase-9 *** knockdown of Smac abrogated cisplatin-induced apoptosis,mitochondrial dysfunction,cytochrome C release,and caspase *** deficiency also reduced the effect of cisplatin on long-term cell viability,and led to increase cisplatin resistance in xenograft tumors in ***,LBW242,a small molecule Smac mimetic,enhanced cisplatin-induced apoptosis and caspase activation,and restored cisplatin sensitivity in Smac-deficient ***:Our data suggested tha

关键词： Apoptosis Smac Esophageal cancer Chemosensitivity

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Liver Xeno-Repopulation with Human Hepatocytes in Fah-/-Rag2-/-Mice after Pharmacological Immunosuppression

Liver Xeno-Repopulation with Human Hepatocytes in Fah-/-Rag2...

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中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Zhiying He Dao Xiang Xiaoyuan Zi Lijian Hui Guangshun Yang Xiaoyan Ding Yiping Hu Xin Wang Haibin Zhang Xin Zhang Dongfu Xie Yixin Chen Kirk J.Wangensteen Stephen C.Ekker Meri Firpo Changcheng Liu Stem Cell Institute University of Minnesota 55455 Minnesota USA Department of Laboratory Medicine and Pathology University of Minnesota 55455 Minnesota USA Department of Genetics Cell Biology and DevelopmentUniversity of Minnesota 55455 Minnesota USA Stem Cell Institute University of Minnesota 55455 Minnesota USA Department of Cell Biology Second Military Medical University 200433 Shanghai China Department of Laboratory Medicine and Pathology University of Minnesota 55455 Minnesota USA Laboratory of Molecular Cell Biology Institute of Biochemistry and Cell Biology Chinese Academy of Sciences 200031Shanghai China Department of Cell BiologySecond Military Medical University 200433 Shanghai China Laboratory of Molecular Cell Biology Institute of Biochemistry and Cell Biology Chinese Academy of Sciences 200031Shanghai China Eastern Hepatobiliary Surgery Hospital Second Military Medical University 200433 Shanghai China Laboratory of Molecular Cell Biology Institute of Biochemistry and Cell Biology Chinese Academy of Sciences 200031Shanghai China

Functional human hepatocytes xeno-engrafted in mouse liver can be used as a model system to study hepatitis virus infection and vaccine *** liver xeno-repopulation has been reported in two kinds of genetically modified mice that have both immune deficiency and liver injury-induced donor hepatocyte selection: the uPA/SCID mice and Fah-/-Rag2-/-Il2rg-/-*** lack of hardy breeding and the overly elaborated technique in these two models may hinder the potential future application of these models to hepatitis virus infection and vaccination *** the transplantation protocol for liver xeno-repopulation from human hepatocytes will increase the model efficiency and application.

关键词： Fah-/-Rag2-/-mice humanized liver pharmacological immunosuppression human hepatocyte transplantation human fetal liver cells

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Phosphorylation and function of eIF4B protein are regulated by PIM kinase family

Phosphorylation and function of eIF4B protein are regulated ...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Jianling Yang,Jun Wang,Guijie Guo,Jilong Chen 中国科学院微生物研究所

Expression of Ab1 genes is associated with a variety of hematopoietic *** mice,the v-Abl oncogene of Abelson virus induces pre-B-cell *** humans,ABL kinase is activated via recombination with other sequences such as the fragment from the BCR locus,an event that plays a key role in the development of chronic myelogenous leukemia(CML) and in some cases of acute lymphocytic *** proteins are non-receptor tyrosine kinases that are normally under tight regulation. However,the oncoproteins v-ABL and BCR-ABL1 are constitutively *** precise mechanisms of how constitutively activated ABL kinases induce cellular transformation remain *** initiation factor 4B(eIF4B) is a multidomain protein with a critical role in the initiation of protein synthesis, and has long been known as a hyperphosphorylated *** has been thought that function of eIF4B can be regulated by ***,in this study we observed the interaction between eIF4B and PIM family members in v-Abl *** PIM family of Ser/Thr kinase,which is consisted of three members:PIM-1,PIM-2,and PIM-3,has been implicated in multiple cellular processes including cellular survival,anti-apoptosis and tumorigenesis,and expression of PIM-1 and PIM-2 is Ab1 kinase-dependent in v-Abl *** identified eIF4B as a Pim kinase *** found that Pim-1,Pim-2 but not Pim-3 can upregulate phosphorylation levels of eIF4B on Ser406 and Ser422,while kinase-dead mutants of Pim-1 or Pim-2 significantly inhibit phosphorylation of ***,in vitro kinase assay revealed that Pim-1 kinase can directly phosphorylate eIF4B on *** expected,we found that Pim-1 kinase directly bound to eIF4B using GST-pull down assay and co-immunoprecipitation ***,levels of eIF4B protein were decreased markedly in v-Abl transformants treated with the Abl kinase inhibitor imatinib,indicating that expression of eIF4B is Abl kinas

关键词： Ab1 oncogene e1F4B PIM phosphorylation signal transduction

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