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Effect of Apolipoprotein A-I on ATP Binding Cassette Transporter A1 Degradation and Cholesterol Efflux in THP-1 Macrophage-derived Foam Cells

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 218-226页

作者： Chao-Ke TANG1,2, Guo-Hua TANG1, Guang-Hui YI1, Zuo WANG1, Lu-Shan LIU1, Shuang WAN1, Zhong-Hua YUAN1, Xiu-Sheng HE1, Jun-Hao YANG1, Chang-Geng RUAN1, and Yong-Zong YANG1* 1Institute of Cardiovascular Disease of Nanhua university, Hengyang 421001, china 2xiangya medical school, central southern university, changsha 410078, china Institute of Cardiovascular Disease of Nanhua university Hengyang 421001 China xiangya medical school Central Southern University Changsha 410078 China

Cholesterol-loaded macrophage foam cells are a central componentof atherosclerotic lesions. ATP binding cassette transporter A1 (ABCA1), the defective molecule inTangier disease, mediates the efflux of phospholipid and cholesterolfrom cells to apolipoprotein A-I (apoA-I), reversing foam cell formation. This study investigated the effect of apoA-I on ABCA1 degradation and cholesterol efflux in THP-1 macrophage- derived foam cells. After exposure of the cultured THP-1 macrophage-derived foam cells to apoA-I for different time, cholesterol efflux, ABCA1 mRNA and protein levels were determined by FJ-2107P type liquid scintillator, RT-PCR and Western blot, respectively. The mean ABCA1 fluorescence intensity on THP-1 macrophage-derived foam cells was detected by flow cytometry. Results showed that apoA-I markedly increased ABCA1-mediated cholesterol efflux from THP-1 macrophage-derived foam cells. This was accompanied by an increase in the content of ABCA1. ApoA-I did not alter ABCA1 mRNA abundance. Significantly, thiol protease inhibitors increased the level of ABCA1 protein and slowed its decay in THP-1 macrophage-derived foam cells, whereas none of the proteosome-specific inhibitor lactacystin, other protease inhibitors, or the lysosomal inhibitor NH4Cl showed such effects. The apoA-I-mediated cellular cholesterol efflux was enhanced by thiol protease inhibitors. Our results suggested that thiol protease inhibitors mightprovide an alternative way to upregulate ABCA1 protein. Thisstrategy is especially appealing since it may mimic the stabilizingeffect of the natural ligands apoA-I.

关键词： ATP binding cassette transporter A1 apoA-I flow cytometry cholesterol lipoproteins

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Identification of Genes Related to Nasopharyngeal Carcinoma with the Help of Pathway-based Networks

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Acta Biochimica et Biophysica Sinica 2006年第12期38卷 900-910页

作者： Hui LI~1 Cai-Ping REN~1* Xiao-Jun TAN~2 Xu-Yu YANG~1 Hong-Bo ZHANG~1 Wen ZHOU~1 Kai-Tai YAO~1 1 Cancer Research Institute,xiangya school of Medicine,central South university,changsha 410078,china 2 Reproductive and Stem Cell Engineering Institute,xiangya school of Medicine,central South university,changsha 410078,china Cancer Research Institute Xiangya School of Medicine Central South UniversityChangsha 410078 China Reproductive and Stem Cell Engineering Institute Xiangya School of Medicine Central South UniversityChangsha 410078 China

cDNA microarray is a powerful tool to analyze simultaneously the expression levels of tens ofthousands of *** with normal nasopharynx (NP) tissues,2210 genes were highly differentiallyexpressed in nasopharyngeal carcinoma (NPC) tissues detected by cDNA *** signal pathwayis widely used to describe the complex relationship between genes,a pathway-based network was con-structed to visualize the connection between the genes obtained from microarray data in this *** the targeted genes that may have more important influence on this gene network with statisticalmethods and found that some genes might have significant influence on this network,especially Ras-relatednuclear protein (RAN),carboxyl ester lipase (CEL),v-rel reticuloendotheliosis viral oncogene homolog A(RELA) *** verify the results from pathway-based selection,reverse transcription-polymerase chainreaction (RT-PCR) and real-time RT-PCR were performed to detect the expression levels of RAN,CEL andRELA genes and it was found that the RAN and CEL genes were significantly up-regulated in more than 80%of NPC *** further elucidate the function of the RAN gene,RAN expression was specifically sup-pressed in a 5-8F NPC cell line by RNA interference (RNAi).As expected,the depletion of RAN couldeffectively block the proliferation of tumor ***,our study may open up a new way to analyze thevast microarray data.

关键词： nasopharyngeal carcinoma microarray real-time PCR RNA interference Ras-related nuclear protein (RAN)

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Polycomb group proteins and their roles in carcinogenesis

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Chinese Science Bulletin 2012年第18期57卷 2259-2264页

作者： XIAO LanBo 1,2,3,TAO YongGuang 1,2,3,LI LiLi 1,2,3,4 & CAO Ya 1,2,31 Key Laboratory of Carcinogenesis of Ministry of Health,xiangya school of Medicine,central South university,changsha 410078,china 2 Key Laboratory of Carcinogenesis and Cancer Invasion of Ministry of Education,xiangya school of Medicine,central South university,changsha 410078,china 3 Cancer Research Institute,xiangya school of Medicine,central South university,changsha 410078,china 4 Cancer Epigenetics Laboratory,State Key Laboratory in Oncology in South china,Chinese university of Hong Kong,Hong Kong 999077,china Key Laboratory of Carcinogenesis of Ministry of Health Xiangya School of Medicine Central South University Changsha China Key Laboratory of Carcinogenesis and Cancer Invasion of Ministry of Education Xiangya School of Medicine Central South University Changsha China Cancer Research Institute Xiangya School of Medicine Central South University Changsha China Cancer Epigenetics Laboratory State Key Laboratory in Oncology in South China Chinese University of Hong Kong Hong Kong China

In the cell nucleus,DNA is wound around histone proteins,which are then packed together to form *** can be chemically tagged by methyl and acetyl *** group (PcG) proteins attach methyl groups to genes,which block their *** is similar to the attachment of methyl groups to gene promoters by DNA methyltransferases (DNMTs).This action is directly linked with tumor initiation and metastasis via the promotion of anti-senescence and anti-apoptosis pathways,and by facilitating epithelial mesenchymal transition (EMT).Cell fate transcriptional factors (CFTFs) and long non-coding RNAs (long ncRNAs) recruit PcG proteins to the promoters of tumor suppressor genes,resulting in epigenetic gene silencing by influencing chromatin structure and DNA ***,PcG proteins are potential diagnostic markers and targets for new chemoprevention and therapeutic strategies.

关键词：组蛋白 DNA甲基转移酶基因启动子癌变非编码RNA PEG 凋亡途径抑癌基因

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miRNA and nasopharyngeal carcinoma

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Chinese Science Bulletin 2011年第8期56卷 722-728页

作者： ZHAO LuQing1,2,3,CHEN Xue1,2,3 & CAO Ya1,2,3 1 Key Laboratory of Carcinogenesis and Invasion,Ministry of Education,changsha 410078,china 2 Key Laboratory of Carcinogenesis,Ministry of Health,changsha 410078,china 3 Cancer Research Institute,xiangya school of Medicine,central South university,changsha 410078,china Key Laboratory of Carcinogenesis and Invasion Ministry of Education Changsha China Key Laboratory of Carcinogenesis Ministry of Health Changsha China Cancer Research Institute Xiangya School of Medicine Central South University Changsha China

MicroRNAs(miRNAs) constitute a family of small non-coding RNA molecules 22-25 nucleotides in *** control the expression of target genes at the post-transcriptional level by inhibiting translation or by degrading target mRNA through binding to complementary sequences in the 3'-untranslated *** studies have demonstrated that miRNAs are intimately involved in processes leading to nasopharyngeal carcinoma,such as the Epstein-Barr virus-encoded latent membrane protein 1 activated signal transduction pathways,gene-regulatory networks,mitosis,tumor angiogenesis,invasion and *** the relationship between miRNAs and the development of nasopharyngeal carcinoma,will further the understanding of this cancer and provide new avenues for diagnosis and treatment.

关键词： miRNAs 鼻咽癌非编码RNA 小分子RNA 信号转导通路潜伏膜蛋白1 基因调控网络肿瘤血管生成

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Schistosoma japonicum: Isolation and Identification of Peptides Mimicking Ferritin Epitopes from Phage Display Library

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 206-210页

作者： Lian-Fei TANG, Xin-Yuan YI*, Xian-Fang ZENG, Lin-Qian WANG, and Shun-Ke ZHANG Department of Parasitology, central South university xiangya school of Medicine, changsha 410078, china Department of Parasitology Central South University Xiangya School of Medicine Changsha 410078 China

In an attempt to isolate and identify the antigenic epitopes on ferritin of Schistosoma japonicum (SjFer) and to test their protective potentiality against Schistosoma japonicum (S.j), polyclonal antisera against SjFer was prepared to screen a 12-mer random peptide library. Three rounds of biopan- ning were performed and resulted in an enrichment. Six peptides selected randomly from the third elute were all found to be positive by evaluating the binding to anti-SjFer sera by ELISA and Western blot- ting. Three amino acid sequences were deduced from the six phage clones by sequencing. When they were used to vaccinate mice, the three peptides could induce significant reduction in adult worms (26.7%, 20.4%, and 25.9%) as well as in liver eggs per gram (LEPG) (40.0%, 38.2%, and 40.8%). This result showed that three mimotopes on SjFer were obtained and they could induce significant protective efficacy against S.j.

关键词： phage displayed peptide library epitope Schistosoma japonicum ferritin

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Sequencing of phosphopeptides sulfonated by 4-sulfophenyl isothiocyanate using post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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Progress in Natural Science:Materials International 2006年第9期16卷 930-935页

作者： TANG Jianguang , WANG Yongjun , ZHANG Hainan, WANG Chunyu, HU Zhiping, XUE Zhigang, XIA Kun3 and SHI Xiaoliu ( Department of Neurology, the Second xiangya Hospital, central South university, changsha 410011, china Department of Digestion, the Second xiangya Hospital, central South university, changsha 410011, china National Key Laboratory of Chinese Genetics, changsha 410078, china) Department of Neurology The Second Xiangya Hospital Central South University Changsha 410011 China Department of Digestion The Second Xiangya Hospital Central South University Changsha 410011 China National Key Laboratory of Chinese Genetics Changsha 410078 China

Phosphorylation/dephosphorylation is probably the most common and important reversible post-translational modification of proteins. Analyzing the functional effects of phosphorylation is helpful for understanding the biological functions of proteins. Identification of the phosphorylation sites of phosphorylated protein is a prerequisite for research on phosphorylation. In this work, an effective and simple method of identification of protein phosphorylation sites has been developed. Phosphopeptides were selectively enriched with immobilized metal affinity chromatography (IMAC) and subsequently chemically modified by 4-sulfophenyI isothiocyanate, and then the chemically modified phosphopeptides were sequenced with post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry for detecting phosphorylation sites. The charge of derivatization by 4-sulfophenyl isothiocyanate introduces a negative sulfonic acid group at the N-terminus of a peptide, and enables the selective detection of only a single series of C-terminal y-type ions. This chemically assisted method greatly simplifies the extremely complex pattern of PSD fragment ions and makes the PSD spectra more easier to be interpreted. The phosphorylation sites of a synthesized model phosphopeptide and human c-myc protein have been successfully identified by this method.

关键词： phosphopeptides sequencing 4-sulfophenyl isothiocyanate post-source decay matrix-assisted laser desorption/ion-ization time-of-flight mass spectrometry.

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Protective effect of ginsenoside Rg1 on glutamate-induced lung injury

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Acta Pharmacologica Sinica 2007年第3期28卷 392-397页

作者： Li SHEN~2 Jian-zhong HAS~2 Chen LI~2 Shao-jie YUE~3 Yong LIU~2 Xiao-qun QIN~2 Hui-jun LIU~2 Zi-qiang LUO~(3,4)2 Departments of Physiology,xiangya medical school and3 Neonatology,xiangya Hospital,central South university,changsha 410078,china Departments of Physiology Xiangya medical school Central South University Changsha China Departments of Neonatology Xiangya Hospital Central South University Changsha China

Aim:To examine the possible protective effect of ginsenoside Rgl,an activecomponent of ginseng,on lung injury caused by glutamate in ***:Thelungs of mice receiving glutamate(0.5 g/kg)and/or ginsenoside Rg1(0.03 g/kg)via intraperitoneal administration were *** indexes of lung wet weight/body weight ratios(LW/BW),lung wet/dry weight ratios(W/D),heart rate(HR),and breathing rate(BR)were *** activity of nitric oxide synthase(NOS),xanthine oxidase(XOD),superoxide dismutase(SOD),catalase(CAT),thecontent of NO,and malondialdehyde in the lung homogenate were ***:Treatment with glutamate for 2 h increased LW/BW,W/D,HR,and *** changes were nearly abolished by pretreatment with ginsenoside Rgl for30 min before glutamate *** analysis of the lung homogenate demon-strated the protective effect as evidenced by the inhibition of NOS(12%)andXOD(50%)inactivity,the enhanced activity of SOD(20%)and CAT(25%).Conclusion:Ginsenoside Rgl has a potential protective role in lung diseasesassociated with glutamate toxicity.

关键词： ginsenoside Rgl glutamate acute lung injury nitrogen monoxide reactive oxygen species

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Genistein stimulates osteoblastic differentiation via p38 MAPK-Cbfa1 pathway in bone marrow culture

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Acta Pharmacologica Sinica 2007年第10期28卷 1597-1602页

作者： Qing-chuan LIAO~2,Zhou-sheng XIAO~(4,5),Yan-fang QIN~3,Hong-hao ZHOU~4~2 Affiliated Children's Hospital,Nanjing medical university,Nanjing 210008,china,~3 Department of Pathology,school of Medicine,Jinanuniversity,Guangzhou 510632,china ~4 Institute of Clinical Pharmacology,central South university,changsha 410078,china Affiliated Children's Hospital Nanjing Medical University Nanjing China Institute of Clinical Pharmacology Central South University Changsha China Department of Pathology School of Medicine Jinan University Guangzhou China

Aim:To test the hypothesis that genistein stimulates the osteoblastic differentia-tion through the p38 mitogen activated protein kinase (MAPK)-core-binding fac-tor 1 (Cbfa1) ***:The activation of p38 MAPK was detected byWestern *** phosphatase (ALP) activity and calcium depositionwere assessed for osteoblastic differentiation of bone marrow-derived mesenchy-mal stem cell (BMSC) *** expression of Cbfa1 was analyzed at both themRNA and protein *** activity of Cbfa1 was detected by electrophoreticmobility shift *** sialoprotein (BSP),ALP,osteocalcin (OC),andosteopontin (OPN) gene transcription were also evaluated by either RT-PCR orNorthern ***:Genistein (0.01-1 μmol/L) dose dependently led tothe rapid and sustained activation of the p38 MAPK pathway in mouse *** with genistein (1 μmol/L) resulted in increased ALP activityand calcium deposition of BMSC cultures as a function of *** alsoenhanced Cbfa1 DNA binding activity and promoted the expressions of Cbfa1itself as well as several Cbfa1-regulated genes,including ALP,BSP,OC,and *** treatment with p38 MAPK inhibitor (SB203580) diminished thegenistein-induced osteoblastic maturation and p38 MAPK-Cbfa1 activation inmouse BMSC ***:These results indicated that genistein couldstimulate the osteoblastic differentiation of BMSC cultures through the p38MAPK-Cbfa1 pathway.

关键词： bone marrow-derived mesenchymal stem cells core-binding factor 1 osteoblastic differentiation genistein mitogen-activated protein kinase

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Vaccination against Schistosoma japonicum Infection by DNA Vaccine Encoding Sj22.7 Antigen

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 27-36页

作者： Gan DAI Shiping WANG Junlong YU Shaorui XU Xianchu PENG Zhuo HE Xueqin LIU Songhua ZHOU and Fen LIU Institute of Schistosomiasis Research.xiangya school of Medicine,central South university,changsha 410078 china Institute of Schistosomiasis Research Xiangya School of Medicine Central South UniversityChangsha 410078 China

To observe the in vitro expression of DNA vaccine pcDNA3-Sj22.7 and its immunologicaleffect in mice,the recombinant plasmid pcDNA3-Sj22.7 was used to transfect HeLa cells with liposome-mediated method and the expression of Sj22.7 mRNA and protein was examined using reversetranscription-polymerase chain reaction, sodium dodecylsulfate-polyacrylamide gel electrophoresis andWestern ***,the ability of pcDNA3-Sj22.7 to protect against Schistosoma japonicum challengeinfections was analyzed according to worm reduction rate and egg reduction rate after vaccination of *** serum levels of specific IgG antibody and T lymphocyte proliferation response were also *** the challenge infection,Sj22.7-driven interferon (IFN)-γand interleukin (IL)-4 was also *** showed that pcDNA3-Sj22.7 could express Sj22.7 mRNA and protein in *** in a worm reduction rate of 29.70%,egg reduction rate of 47.25% (liver) and 51.73% (intestine),and egg reduction rate of 25.90% (eggs per female),suggesting induction of significant anti-fecundityin the pcDNA3-Sj22.7 group. Enzyme-linked immunosorbent assay and Western blot analysis indicatedthat immunized mice generated specific IgG against Sj22.7.T lymphocytes from mice immunized withpcDNA3-Sj22.7 showed a significant proliferation response to *** culture of spleen cells showedthat secretion of IFN-γ increased but IL-4 decreased. The results indicate that DNA vaccination bypcDNA3-Sj22.7 is sufficient to elicit significant levels of protective immunity against *** *** DNA vaccine could induce significant cellular and humoral immune response,and display predominantT helper cell type 1 type immune responses,which contribute to the protective immunity against challengeinfection in mice.

关键词： response Schistosoma japonicum DNA vaccine Sj22.7 gene expression immune protection immune

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Genetic and Epigenetic Alterations of DLC-1,a Candidate Tumor Suppressor Gene,in Nasopharyngeal Carcinoma

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 349-355页

作者： Dan PENG Cai-Ping REN Hong-Mei YI Liang ZHOU Xu-Yu YANG Hui LI Kai-Tai YAO Cancer Research Institute,xiangya school of Medicine,central South university,changsha 410078,china Cancer Research Institute Xiangya School of Medicine Central South UniversityChangsha 410078 China

The DLC-1 gene,located at the human chromosome region 8p22,behaves like a tumorsuppressor gene and is frequently deleted in diverse *** deletion of 8p22 is not an uncommon eventin nasopharyngeal carcinoma(NPC),therefore we explored the expression levels of the DLC-1 gene in NPCsand NPC cell lines by reverse transcription-polymerase chain *** results showed the mRNA levelof DLC-1 was *** identify the mechanism of DLC-1 downregulation in NPC,we investigatedthe methylation status of the DLC-1 gene using methylation-specific PCR,and found that 79%(31 of 39)ofthe NPC tissues and two DLC-1 nonexpressing NPC cell lines,6-10B and 5-8F,were methylated in theDLC-1 CpG *** PCR was also carried out,and loss of heterozygosity was found at fourmicrosatellite sites(D8S552,D8S 1754,D8S 1790 and D8S549)covering the whole DLC-1 gene with ratiosof 33%(4 of 12 informative cases),18%(2 of 11),5%(1 of 18),and 25%(3 of 12),***,our results suggest that DLC-1 might be an NPC-related tumor suppressor gene affected by aberrantpromoter methylation and gene deletion.

关键词： DLC-I nasopharyngeal carcinoma hypermethylation loss of heterozygosity

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