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Adenovirus-mediated Expression of both Antisense Ornithine Decarboxylase and S-adenosylmethionine Decarboxylase Inhibits Lung Cancer Cell Growth

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Acta Biochimica et Biophysica Sinica 2007年第6期39卷 423-430页

作者： Hui TIAN~(1*), Xianxi LIU~2. Bing ZHANG~2. Qifeng SUN~1, and Dongfeng SUN~1 1 department of Thoracic Surgery, Qi Lu Hospital, shandong University {2. 2.0012. China 2.department of medicine, medical molecular biology experimental Center, shandong University {2. 2.0012.China department of Thoracic Surgery Qi Lu Hospital Shandong UniversityJinan 250012 China department of medicine Medical Molecular Biology Experimental Center Shandong UniversityJinan 250012 China

Polyamine biosynthesis is controlled primarily by ornithine decarboxylase （ODC） and Sadenosylmethionine decarboxylase （AdoMetDC）. Antisense sequences of ODC and AdoMetDC genes were cloned into an adenoviral vector （named Ad-ODC-AdoMetDCas）. To evaluate the effects of recombinant adenovirus Ad-ODC-AdoMetDCas that can simultaneously express both antisense ODC and AdoMetDC, the human lung cancer cell line A-549 was infected with Ad-ODC-AdoMetDCas or the control vector. Viable cell counting, determination of polyamine concentrations, cell cycle analysis, and Matrigel invasion assays were carried out to assess the properties of tumor growth and invasiveness. Our study showed that adenovirus-mediated antisense ODC and AdoMetDC expression inhibits tumor cell growth through blocking the polyamine synthesis pathway. Tumor cells were arrested at the G1phase after gene transfer and the invasiveness was reduced. It suggested that the recombinant adenovirus Ad-ODC-AdoMetDCas might be a new anticancer reagent in the treatment of lung cancers.

关键词： ornithine decarboxylase S-adenosylmethionine decarboxylase polyamine lung cancer genetherapy

Gene Expression of Ornithine Decarboxylase in Lung Cancers and Its Clinical Significance

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Acta Biochimica et Biophysica Sinica 2006年第9期38卷 639-645页

作者： Hui TIAN~1* Qing HUANG~1 Lm LI~1 Xian-Xi LIU~2.Yan ZHANG~2. 1 department of Thoracic Surgery,shandong University Qi Lu Hospital,{2. 2.0012.China 2.department of medicine,medical molecular biology experimental Center,shandong University,{2. 2.0012.China department of Thoracic Surgery Shandong University Qi Lu HospitalJinan 250012 China department of medicine Medical Molecular Biology Experimental Center Shandong UniversityJinan 250012 China

Lung cancer is one of the most lethal cancers in China because of its high incidence and *** decarboxylase （ODC）,an important enzyme in polyamine biosynthesis,is increased incancer *** chemotherapeutic agents aimed at reducing ODC expression show inhibitory effects oncancer cell growth,so ODC can be useful in gene diagnosis and gene therapy of *** this study,weexamined the effect of antisense ODC on lung cancer cells.A-549 cells were infected with rAd-ODC/Ex3as,a recombinant adenovirus containing the cytomegalovirus promoter,green fluorescent protein gene and 12.bp antisense *** cell cycle was evaluated by flow cytometry.A nude mouse xenograft model wasused in the tumorigenicity *** transcription-polymerase chain reaction,Western blot and immuno-histochernistry were used to study the expressions of ODC on lung *** was found that the growth ofcells infected with rAd-ODC/Ex3as was substantially inhibited and cells were arrested at *** with rAd-ODC/Ex3as can suppress tumor formation in a nude mouse xenograft *** of ODC mRNA and ODC protein levels in lung cancer tissues was significantly higher than thatin normal tissues （P<0.05）,which correlated significantly with the stage of lung cancer （P<0.05）.This studysuggested that rAd-ODC/Ex3as has antitumor activity in human lung cancer *** ODC gene might playan important role in lung cancer and the overexpression of ODC might be related to the occurrence anddevelopment of lung cancer.

关键词： polyamine biosynthesis lung neoplasm ornithine decarboxylase A-549 cell line

Effects of Raloxifene on Caveolin-1 mRNA and Protein Expressions in Vascular Smooth Muscle Cells

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 747-752页

作者： Fa-Lin YANG~(1#*) Hong HE~(2.) Xian-Xi LIU~1* Bing TU~1 Xian-Wei ZENG~3 Ji-Xin SU~1 Xin WANG~2.Qin HU~2. 1 Institute of Biochemistry and molecular biology,{2. College,shandong University,jinan 2.0012.China 2.department of Cardiology,Qilu Hospital,shandong University,jinan 2.0012.China 3 department of Clinical medicine,Weifang {2. College,Weifang 2.0041,China Institute of Biochemistry and molecular biology Medical College Shandong UniversityJinan 250012 China department of Cardiology Qilu Hospital Shandong UniversityJinan 250012 China department of Clinical medicine Weifang Medical CollegeWeifang 260041 China

Caveolin-1 is regulated by estrogen in vascular smooth muscle ***,a selectiveestrogen receptor modulator that possibly has cardioprotective properties without an increased risk of canceror other side effects of estrogen,may be used in women with risk of coronary artery ***,therelationship between raloxifene and caveolin-1 is still ***,this study was designed to seewhether raloxifene regulates caveolin-1 expression and if so,whether such regulation is mediated by *** aortic smooth muscle cells were cultured in the absence or presence of raloxifene (10-8to10-6M) for 12.or 2. *** mRNA and protein levels of caveolin-1 were increased significantly after 2. htreatment with *** increases were inhibited by estrogen receptor antagonist ICI 182.80 (10-5M).Results of this study suggest that raloxifene stimulates caveolin-1 transcription and translation throughestrogen receptor mediated mechanisms.

关键词： raloxifene caveolin-1 estrogen receptor vascular smooth muscle cell

Adenovirus-mediated expression of both antisense ODC and AdoMetDC inhibited colorectal cancer cell growth in vitro

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Acta Pharmacologica Sinica 2006年第3期27卷 353-359页

作者： Bing ZHANG~2.Xian-xi LIU~(2.5) Yan ZHANG~2.Chun-ying JIANG~3 Qing-shan TENG~4 Hai-yan HU~2.Wei WANG~2.Lei GONG~2.~2.nstitute of Biochemistry and molecular biology,School of {2..shandong University,jinan 2.0012.China ~3department of Coloproctology,Affiliated Hospital of shandong University of Traditional Chinese {2.,jinan 2.0014,China,~4department of Neurosurgery.Cleveland Clinic Foundation,Cleveland,OH 44195,USA Institute of Biochemistry and molecular biology School of Medicine Shandong University Jinan China department of Colo-proctology Affiliated Hospital of Shandong University of Traditional Chinese Medicine Jinan China department of Neurosurgery Cleveland Clinic Foundation Cleveland USA

Aim:To construct a recombinant adenovirus that can simultaneously expressboth antisense ornithine decarboxylase(ODC)and adenosylmethionine decar-boxylase(AdoMetDC)and detect its inhibitory effect on the intracellular polyaminepool and colorectal cancer cell ***:A 2.5-bp cDNA of AdoMetDCwas reverse-inserted into recombinant pAdTrack-ODCas vectors and recombinedwith pAdEasy-1 vectors in AdEasy-1 *** clones were selected andtransfected into the packaging cell HEK2.3 after they were linearized by *** fluorescent protein expression was used to monitor the process of adenovi-rus *** ODC and AdoMetDC protein levels were identified by west-ern blotting,and intracellular polyamine content was detected by reverse-phasehigh performance liquid chromatography.A viable cell count was used to deter-mine the growth of HT-2. cells with or without exogenous ***:Sequencing confirmed that AdoMetDC cDNA was successfully ligated into thepAdTrack-ODCas *** expression in 2.3 cells during virus packing andamplification was observed by fluorescence *** blotting dem-onstrated that both ODC and AdoMetDC were downregulated by Ad-ODC-AdoMetDCas,and consequently 3 kinds of polyamine(putrescine,spermidineand spermine)were reduced to very low ***-2. cell growth was signifi-cantly inhibited as compared with control conditions,and growth arrest was notreversed by exogenous ***:The successfully constructedrecombinant adenovirus,Ad-ODC-AdoMetDCas,blocked polyamine synthesisand has therapeutic potential for treating colorectal cancer in vitro.

关键词： polyamine ornithine decarboxylase adenosylmethionine decarboxylase adenovirus colorectal cancer

Recurrence spectra of He atoms in strong external fields

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Science China(Physics,Mechanics & Astronomy) 2004年第4期47卷 477-484页

作者： LIN Shenglu1, LI Hongyun1, WANG Dehua2. ZHAO Wenli1 & GAO Feng1 1. department of Physics, shandong Normal University, {2. 2.0014, China 2. department of Mathematics and Physics, shandong Institute of Architecture and Engineering, {2. 2.0014, China department of Physics Shandong Normal University Jinan China department of Mathematics and Physics Shandong Institute of Architecture and Engineering Jinan China

By employing a model potential including the electron exchange energy, we extend the semiclassical closed orbit theory to study the multielectron atoms. Using special region-splitting consistent and iterative method, we figure out the closed orbits in the corresponding classical system and calculate the recurrence spectra of triplet helium atoms in parallel electric and magnetic fields at scaled energy ε=?0.03,n≈40,n=0. The core-scattering effects have been taken into account, which lead to more peaks in the spectra. It has also been confirmed by means of the direct comparison between the spectral portrait in such a plot and those of hydrogen case. In order to compare the theoretic results with experiment, we investigate the closed orbits and recurrence spectra of helium atoms for the similar exchange potential but applied only by single electric field at scaled energy ε=?2.7 case. The spectra are in good agreement with the experimental observation. We conclude that our model is correct and it is necessary to consider the exchange effect for determining the photoabsorption spectra of multielectron atoms in strong external fields.

关键词： exchange potential, recurrence spectra, semiclassical closed orbit theory, core-scattering.

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Morphology and electrical conductivity of PACS

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Journal of Environmental Sciences 2000年第3期12卷 319-324页

作者： GAO Bao\|yu\+1, LIU Zong\|gang\+2. YUE Qin\|yan\+3, ZHAO Hua\|zhang\+3 (1. department of Environmental Engineering, shandong University, {2. 2.0100, China 2. Environmental Monitoring Station of Huaiyin, {2. 2.002., China 3. department of Environ 山东大学山东济南 2 50 1 0 0 山东济南市槐荫区环境监测山东济南 2 50 0 2 2

Polyaluminium chlorides with sulfate ion(PACS) were prepared by using AlCl 3·6H 2., Al(SO 4) 3·18H 2. and Na 2.O 3 as raw materials. The effects of basicity ( r ), Al 3+ /SO 4 2. molar ratio and aging time on the morphology of PACS were observed by transmission electrical microscope. The influence of aging time on charge neutralization and coagulation effect of PACS was studied. The effects of basicity ( r ), Al 3+ /SO 4 2. molar ratio and aging time on the electrical conductivity of PACS solution were also investigated. The experimental results show that the degree of polymerization of polyaluminium chloride(PAC) increases when SO 4 2. ion is added. The higher the basicity( r ) and the longer the aging time, the larger the size of polymer PACS. The ability of PACS neutralizing the charge on Kaolinite decreases with the increase of aging time. The electrical conductivity of PACS solution(the concentration of Al 3+ ion is 0.18 mol/L) with different aging time is the function of the basicity and Al 3+ /SO 4 2. molar ratios and has its maximum at r =0.5 and Al 3+ /SO 4 2. =12.

关键词： coagulant PACS morphology transmission electron microscope electrical conductivity coagulation

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The Effect of Anti-Human CD134 Monoclonal Antibody on Phytohemagglutinin-Induced mRNA Expression of Perforin in Peripheral Blood Mononuclear Cells

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Cellular & molecular Immunology 2005年第6期2卷 467-471页

作者： Ming Li~1 Yuanchao Zhang~(1,2. ~1department of Rheumatology and Immunology,shandong Provincial Hospital,shandong University,jinan 2.002.,China ~2.0} of Rheumatology and Immunology,shandong Provincial Hospital,shandong University,jinan 2.002.,shandong,China. department of Rheumatology and Immunology Shandong Provincial Hospital Shandong University Jinan 250021 China

CD134, a member of the tumor necrosis factor receptor superfamily, plays a crucial role in T cell survival. In this study, peripheral blood mononuclear cells （PBMCs） stimulated with phytohemagglutinin （PHA, 50 μg/ml） were treated by CD134 mAb （1μg/ml, 5 μg/ml, 10 μg/ml） for 6 h, 12.h, 2. h and 48 h. The level of perforin mRNA was measured by reverse transcription-polymerase chain reaction （RT-PCR） technique. Our data showed that the expression of perforin mRNA in PBMCs was down-regulated by CD134 mAb in a dose-dependent manner in range of 1 μg/ml to 5 μg/ml and dropped down to its minimum on 2. h （p 〈 0.05）. The level of perforin mRNA reached a plateau when the concentration of CD134 mAb exceeded 5 μg/ml. In conclusion, CD134 mAb can inhibit perforin expression, which may enhance the ability of T cells for survival. Cellular ＆ molecular Immunology. 2.05;2.6）： 467-471.

关键词： CD134 perforin phytohemagglutinin

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Cloning,Sequencing and Expression Analysis of the First Cellulase Gene Encoding Cellobiohydrolase 1 from a Cold-adaptive Penicillium chrysogenum FS010

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Acta Biochimica et Biophysica Sinica 2007年第2期39卷 101-107页

作者： Yunhua HOU~(1,2. Tianhong WANG~1 Hao LONG~1 and Huiynan ZHU~(1,2. ~1State Key Laboratory of Microbial Technology,shandong University,jinan 2.0100,China ~2.2. of Food and Biologic Engineering,shandong Institute of Light Industry,jinan 2.032.,China State Key Laboratory of Microbial Technology Shandong University Jinan 250100 China department of Food and Biologic Engineering Shandong Institute of Light Industry Jinan 250323 China

A cellobiohydrolase 1 gene (cbhl) was cloned from Penicillium chrysogenum FS010 by amodified thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR).DNA sequencing showsthat cbhl has an open reading frame of 1590 bp, encoding a putative protein of 52. amino acid *** amino acid sequence revealed that CBHI has a modular structure with a predicted molecular massof 56 kDa and consists of a fungal type carbohydrate binding module separated from a catalytic domain bya threonine rich linker *** putative gene product is homologous to fungal cellobiohydrolases inFamily 7 of the glycosyl hydrolases.A novel cbhl promoter (1.3 kb) was also cloned and sequenced,whichcontains seven putative binding sites (5'-SYGGRG-3') for the carbon catabolite repressor *** ofvarious carbon sources to the cbhl transcription of *** was examined by Northern analysis,suggesting that the expression of cbhl is regulated at transcriptional *** cbhl gene in cold-adaptivefungus *** was expressed as an active enzyme in Saccharomyces cerevisiae *** recombinantCBHI accumulated intracellularly and could not be secreted into the medium.

关键词： Penicillum chrysogenum cellobiohydrolase TAIL-PCR promoter of cbh1

Curcumin downregulates homeobox gene NKX3.1 in prostate cancer cell LNCaP

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Acta Pharmacologica Sinica 2007年第3期28卷 423-430页

作者： Hui-na ZHANG~(2.4) Chun-xiao YU~(2.4) Peng-ju ZHANG~2.Wei-wen CHEN~2.An-li JIANG~(2.5) Feng KONG~2.Jing-ti DENG~2.Jian-ye ZHANG~(2.5) Charles YF YOUNG~3 ~2.nstitute of Biochemistry and molecular biology,{2. School,shandong University,Ji-nan 2.0012.China ~3department of Urology Research,Mayo Clinic,Rochester,MN 55905,USA Institute of Biochemistry and molecular biology Medical School Shandong University Ji-nan China department of Urology Research Mayo Clinic Rochester USA

Aim:To elucidate the effect and the mechanisms of curcumin on the expression ofthe human homeobox gene NKX3.1 in the prostate cancer cell ***:The expression change of NKX3.1 in cells incubated with varying concentrationsof curcumin was observed by Western blotting and RT-PCR.A dual luciferasereporter assay was used to test the effect of curcumin on the activity of the NKX3.11040 bp ***-treated cells disposed to a designated amount ofandrogen analog R1881 and the androgen receptor(AR)antagonist flutamide,then the expression of NKX3.1 or the activity of the NKX3.1 promoter were inves-tigated by Western blotting or reporter gene assay,***,Westernblotting and electrophoretic mobility shift assay were performed to demonstratethe effect of curcumin on the expression of AR and its binding activity to theandrogen response element(ARE).Results:Curcumin downregulated the ex-pression of NKX3.1 and the activity of the NKX3.1 1040 bp promoter in LNCaPcells.R1881 increased the expression of NKX3.1,and the AR antagonist flutamidedecreased the expression of NKX3.1 in LNCaP cells,while curcumin could inhibitandrogen-AR mediated induction of NKX3.1 *** decreased theexpression of AR and the binding activity to ARE ***:Curcumincould downregulate NKX3.1 expression in LNCaP *** could also inhibit theandrogen-AR mediated induction of NKX3.1 expression by downregulating ARexpression and blocking its DNA binding activity.

关键词： curcumin prostate cancer cell NKX3 1 androgen receptor androgen responsive element