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Efficacy of sequential treatment of HCT116 colon cancer monolayers and xenografts with docetaxel,flavopiridol,and 5-fluorouracil

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Acta Pharmacologica Sinica 2006年第10期27卷 1375-1381页

作者： Jun GUO~2 an-wu ZHOU~2 Yu-cai FU~3 Udit N VERMA~2 Debu TRIPATHY~2 Eugene P FRENKEL~(2,5) Carlos R BECERRA~4 ~2Harold C.Simmons Cancer Center,Division of Hematology-Oncology,Department of Medicine,university of Texas Southwestern medical Centel.Dallas,Texas 75390,USA ~3laboratory of cell senescence,shantou university medical college,shantou 515041,china ~4Sammons Cancer Center,Baylor university medical Center,TOPA-US Oncology,Dallas,Texas 75246,USA Division of Hematology-Oncology Department of Medicine Harold C. Simmons Cancer Center University of Texas Southwestern Medical Center Dallas USA laboratory of cell senescence Shantou University Medical College Shantou China Sammons Cancer Center Baylor University Medical Center TOPA-US Oncology Dallas USA

Aim:Clinical treatment of solid tumors with docetaxel,flavopiridol,or 5-fluorou-racil(5-FU)often encounters undesirable side effects and drug *** aims to evaluate the potential role of combination therapy with docetaxel,flavopiridol,or 5-FU in modulating chemosensitivity and better understand howthey might be used ***:HCT116 colon cancer cells were treatedwith docetaxel,flavopiridol,and 5-FU in several different administrative sche-dules in vitro,either sequentially or *** survival was measuredby MTT *** activity of caspase-3 was determined by caspase-3 assaysand the soft agar colony assay was used to test the colony formation of HCT116cells in soft *** also established xenograft models to extend in vitro obser-vations to an in vivo ***:The maximum cytotoxicity was found whenhuman colon cancer HCT116 cells were treated with docetaxel for 1 h followed byflavopiridol for 24 h and 5-FU for another 24 *** sequential combinationtherapy not only inhibits tumor cell growth more strongly compared to othercombination therapies but also significantly reduces colony formation in soft agarand augments apoptosis of HCT116 *** of docetaxel followed 1 hlater by flavopiridol,followed 24 h later by 5-FU in xenograft models,also resultedin delayed tumor growth and higher survival ***:These resultshighlight the importance of an administrative schedule when combining docetaxelwith flavopiridol and 5-FU,providing a rationale explanation for its developmentin clinical trials.

关键词： combination drug therapy docetaxel flavopiridol fluorouracil colonic neoplasms

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Effect of a cancer vaccine prepared by fusions of hepatocarcinoma cells with dendritic cells

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World Journal of Gastroenterology 2001年第5期7卷 690-694页

作者： Juan Zhang~1 Jin-Kun Zhang~2 Shao-Hong Zhuo~3 Hai-Bin Chen~2 1 Clinical laboratory,The First Affiliated Hospital of shantou university medical college,shantou 515041,Guangdong Province,china2 Cancer Pathology laboratory,shantou university medical college,shantou 515031,Guangdong Province,china3 Department of Gastroenterology,Third Municipal Hospital of shantou,shantou 515073,Guangdong Province,china Clinical laboratory The First Affiliated Hospital of Shantou University Medical College Shantou 515041 Guangdong Province China. juanzh005@***

AIM: To prepare a cancer vaccine (H(22)-DC) expressing high levels of costimulatory molecules based on fusions of hepatocarcinoma cells (H(22)) with dendritic cells (DC) of mice and to analyze the biological characteristics and induction of specific CTL activity of H(22)-DC. METHODS: DCs were isolated from murine spleen by metrizamide density gradient centrifugation, purified based on its characteristics of semi-adhesion to culture plates and FcR-,and were cultured in the medium containing GM-CSF and IL-4. A large number of DC were harvested. DCs were then fused with H(22) cells by PEG and the fusion cells were marked with CD11c MicroBeads. The H(22)-DC was sorted with Mimi MACS sorter. The techniques of cell culture, immunocytochemistry and light microscopy were also used to test the characteristics of growth and morphology of H(22)-DC in vitro. As the immunogen, H(22)-DC was inoculated subcutaneously into the right armpit of BALB/C mice, and their tumorigenicity in vivo was observed. MTT was used to test the CTL activity of murine spleen in vivo. RESULTS: DC cells isolated and generated were CD11c+ cells with irregular shape, and highly expressed CD80, CD86 and CD54 molecules. H22 cells were CD11c- cells with spherical shape and bigger volume, and did not express CD80, CD86 and CD54 molecules.H(22)-DC was CD11c+ cells with bigger volume, being spherical, flat or irregular in shape, and highly expressed CD80, CD86 and CD54 molecules, too. H(22)-DC was able to divide and proliferate in vitro, but its activity of proliferation was significantly decreased as compared with H(22) cells and its growth curve was flatter than H(22) cells. After subcutaneous inoculation over 60 days, H(22)-DC showed no tumorigenecity in mice, which was significantly different from control groups (Pcells in mice implanted with fresh H(22)-DC was significantly higher than control groups (P < 0.01). CONCLUSION: H(22)-DC could significantly st

关键词： Cancer Vaccines Animals Antigens, CD Antigens, CD80 Antigens, CD86 cell Fusion Dendritic cells Integrin alphaXbeta2 Intercellular Adhesion Molecule-1 Liver Neoplasms, Experimental control Male Membrane Glycoproteins Mice Mice, Inbred BALB C Research Support, Non-U.S. Gov't Spleen

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Inhibition of tryptase and chymase induced nucleated cell infiltration by proteinase inhibitors

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Acta Pharmacologica Sinica 2004年第12期25卷 111-118页

作者： Shao-heng HE2,3, Han-qiu CHEN, Jian ZHENG Allergy& Inflammation Research Institute, shantou university medical college, shantou 515031, china 2Immunopharmacology Group, university of Southampton, UK Immunopharmacology Group University of Southampton Allergy & Inflammation Research Institute Shantou University Medical College Shantou 515031

AIM: To investigate the ability of proteinase inhibitors to modulate nucleated cell infiltration into the peritoneum of mice induced by tryptase and chymase. METHODS: Human lung tryptase and skin chymase were purified by a similar procedure involving high salt extraction, heparinagarose affinity chromatography followedby S-200 Sephacryl gel filtration chromatography. The actions of proteinase inhibitors on tryptase and chymase induced nucleated cell accumulation were examined with a mouse peritoneum model. RESULTS: A selective chymase inhibitor Z-Ile-Glu- Pro-Phe-CO2Me (ZIGPPF) was able to inhibit approximately 90 % neutrophil, 73 % eosinophil, 87 % lymphocyte and 60 % macrophage accumulation induced by chymase at 16 h following injection. Soy bean trypsin inhibitor (SBTI), chymostatin, and α1-antitrypsin showed slightly less potency than ZIGPPF in inhibition of the actions of chymase. While all tryptase inhibitors tested were able to inhibit neutrophil, eosinophil, and macrophage accumu- lation provoked by tryptase at 16 h following injection, only leupeptin, APC366, and aprotinin were capable of inhibiting tryptase induced lymphocyte accumulation. The inhibitiors of tryptase tested were also able to inhibit tryptase induced neutrophil and eosinophil accumulation at 6 h following injection. When being injected alone, all inhibitors of chymase and tryptase at the concentrations tested by themselves had no significant effect on the accumulation of nucleated cells in the peritoneum of mice at both 6 h and 16 h. CONCLUSION: Proteinase inhibitors significantly inhibited tryptase and chymase-induced nucleated cell accumulation in vivo, and therefore they are likely to be developed as a novel class of anti-inflammatory drugs.

关键词： tryptase chymase nucleated cells proteinase inhibitor

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Liposome-mediated IL-28 and IL-29 expression in A549 cells and antiviral effect of IL-28 and IL-29 on WISH cells

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Acta Pharmacologica Sinica 2006年第4期27卷 453-459页

作者： Ming-cai LI Hao-yang WANG Hai-yan WANG Tao LI Shao-heng HE~2 Allergy and Inflammation Research Institute.shantou university medical college.shantou 515041,china Allergy and Inflammation Research Institute Shantou University Medical College Shantou China

Aim:To construct the recombinant vectors carrying interleukin(IL)-28A,IL-28Band IL-29 cDNAs and express them in human A549 cells,and analyze their antivi-ral activity in vesicular stomatitis virus(VSV)-infected human immortalized am-nion epithelial cell line(WISH cells).Methods:Total cell RNA was extracted fromhuman peripheral blood mononuclear cells(PBMC)activated with poly I:*** encoding human IL-28A,IL-28B,and IL-29 were amplified by reverse-transcription polymerase chain reaction(RT-PCR)and inserted into pcDNA3.1/V5-His-TOPO *** recombinant vectors were transfected into humanA549 cells by a liposome-mediated gene transfer *** RT-PCR and Western blotting were used to detect the mRNA and protein expressionof IL-28A,IL-28B,and *** antiviral activity of IL-28A,IL-28B,and IL-29was determined by a cytopathic effect reduction assay on WISH cells using VSVas a challenge ***:The DNA sequences of the inserts were identical tothe published sequences encoding IL-28A,IL-28B,and IL-29 in *** wasdemonstrated that IL-28A,IL-28B,and IL-29 genes were markedly transcribed intransfected *** of all 3 interleukins in A549 cells was confirmed byWestern blot ***-28 and IL-29 expressed by A549 cells,like interferon(IFN)α-2b,were able to protect WISH cells against VSV ***:IL-28 and IL-29 cDNAs were successfully cloned and expressed in eukaryoticcells via transfection with pcDNA3.1/V5-His-TOPO-IL-28/*** withthis vector produced a specific antiviral activity similar to that of IFN-α,which willprovide a new tool for the functional study of these cytokines in humans.

关键词： human interleukin-28A protein human interleukin-28B protein human interleukin-29 protein gene expression liposomes antiyiral agents

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Activation of human tonsil and skin mast cells by agonists of proteinase activated receptor-2

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Acta Pharmacologica Sinica 2005年第5期26卷 568-574页

作者： Shao-heng HE~2,Hua XIE,Yi-ling FUAllergy & Inflammation Research Institute,shantou university medical college.shantou 515031,china Allergy and Inflammation Research Institute Shantou University Medical College Shantou 515031 China. shoahenghe@***

Aim:To investigate the effects of the agonists of proteinase activated receptor(PAR)-2,and histamine on degranulation of human mast ***:Humanmast cells were enzymatically dispersed from tonsil and skin *** dis-persed cells were then cultured with various stimuli,and tryptase and histaminelevels in cell supernatants collected from challenge tubes were ***:PAR-2 agonist peptide SLIGKV provoked a dose-dependent release of histaminefrom skin mast *** also induced tryptase release from tonsil mast ***-LIGRLO appeared less potent than SLIGKV in induction of release of histamineand *** was able to induce a"bell"shape increase in tryptaserelease from tonsil mast *** was also able to induce a dose-dependent releaseof histamine from both tonsil and skin mast *** actions of trypsin on mastcells were inhibited by soy bean trypsin inhibitor(SBTI)or α1-antitrypsin(α1-AT).Time course study revealed that both stimulated tryptase or histamine releaseinitiated within 10 s and reached their peak release between 4 and 6 ***-ment of cells with metabolic inhibitors or pertussis toxin reduced the ability ofmast cells to release tryptase or ***:It was demonstrated thatthe in vitro tryptase release properties of human tonsil and skin mast cells sug-gested a novel type of mast cell *** activation of mast cells byPAR-2 agonists indicated a self-amplification mechanism of mast cell degranulation.

关键词： tryptase histamine mast cells proteinase activated receptor-2 anti-IgE tonsil skin

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Effects of N-n-butyl Haloperidol Iodide on Myocardial Ischemia/ReperfusionInjury and Egr-1 Expression in Rat

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Acta Biochimica et Biophysica Sinica 2006年第6期38卷 435-441页

作者： Yan-Mei ZHANG~1,Gang-Gang SHI~(1*),Zhao TANG~1,Jin-Hong ZHENG~1,Wei-Qiu LI~2,Fu-Xiao GUO~1,and Qiang-Yong JIA~1~1 Department of Pharmacology and~2 Central laboratory,shantou university medical college,shantou 515041,china Department of Pharmacology Shantou University Medical CollegeShantou 515041 China Central laboratory Shantou University Medical CollegeShantou 515041 China

We have previously shown that N-n-butyl haloperidol iodide (F) derived from haloperidolreduces ischemia/reperfusion-induced myocardial injury by blocking intracellular *** studytested the hypothesis that cardio-protection with F2 is associated with an attenuation in the expression ofearly growth response gene 1 (Egr-l).In an in vivo rat model of 60 rain coronary occlusion followed by180 rain of reperfusion,treatment with Fsignificantly reduced myocardial injury evidenced by the reductionin release of plasma creatine kinase,myocardial creatine kinase isoenzyme and lactate *** neonatal rat cardiomyocytes of hypoxia for 3 h and reoxygenation for 1 h,F2 treatment attenuatednecrotic and apoptotic cell death,as demonstrated by electron *** with cardio-protectionby F,the increased expression levels of Egr-l mRNA and protein were significantly reduced in myocardialtissue and cultured cardiomyocytes as detected by reverse transcription-polymerase chain reaction,immunohistochemistry and *** conclusion,these results suggest that the protectiveeffect of Fon ischemia/reperfusion-or hypoxia/reoxygenation-induced myocardial injury might be partlymediated by downregulating Egr-l expression.

关键词： N-n-butyl haloperidol iodide ischemia/reperfusion injury early growth response gene-1 myocardium cardiomyocyte

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Inhibition of histamine release from human mast cells by natural chymase inhibitors

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Acta Pharmacologica Sinica 2004年第6期25卷 120-124页

作者： Shao-heng HE2,Hua XIE, Xiao-jun ZHANG,Xian-jie WANG Allergy & Inflammation Research Institute, shantou university medical college, shantou 515031, china Allergy & Inflammation Research Institute Shantou University Medical College 上海 515031

AIM: To investigate the ability of natural chymase inhibitors to modulate histamine release from human mast cells. METHODS: Enzymatically dispersed cells from human lung, tonsil, and skin were challenged with anti-IgE or calcium ionphore A23187 in the absence or presence of the natural chymase inhibitors secretory leukocyte pro- tease inhibitor (SLPI) and α1-antitrypsin, then histamine release was determined. RESULTS: IgE-dependent hista- mine release from lung, tonsil, and skin mast cells were inhibited by up to 70%, 61%, and 62%, respectively following incubation with α1-antitrypsin (5000 nmol/L).SLPI 5000 nmol/L was also able to inhibit anti-IgE- dependent histamine released from lung, tonsil and skin mast cells by up to approximately 72%, 67%, and 58%, respectively. While neither α1-antitrypsin nor SLPI by themselves altered histamine release from lung, tonsil and skin mast cells, they were able to inhibit calcium ionophore-induced histamine release from lung and tonsil mast cells. CONCLUSION: Both α1-antitrypsinand SLPIcould potently inhibit IgE-dependent and calcium ionophore- induced histamine release from dispersed human lung, tonsil, and skin mast cells in a concentration-dependent manner, which suggested that they were likely to play a protective role in mast cell associated diseases including allergy.

关键词： chymase mast cells immunoglobulin E histamine release calcium ionophore secretory leukocyte protease inhibitor α1-antitrypsin allergy

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Molecular mechanisms of adenosine-induced apoptosis in human HepG2 cells

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Acta Pharmacologica Sinica 2006年第4期27卷 477-484页

作者： Ling-fei WU~(2,4) Guo-ping LI~2 Jia-lin FENG~3 Ze-jin PU~2 ~2Department of Gastroenterology:~3Department of Information,Second Affiliated Hospital,shantou university medical college,shantou 515041.china Department of Gastroenterology Second Affiliated Hospital Shantou University Medical College Shantou China Department of Information Second Affiliated Hospital Shantou University Medical College Shantou China

Aim:To investigate effects of adenosine on cell proliferation and apoptosis inhuman HepG2 ***:HepG2 cells were incubated in the presence ofadenosine(0.1-5 mmol/L)for 12-48 h,and the effect of adenosine on cell prolifera-tion was evaluated by using 3-(4,5-dimethyl-2-yl)-2,5-diphenyl-2H-tetrazolium bro-mide(MTT)*** 33342 fluorescent staining,dUTP-fluoresceinisothiocyanate(FITC)fluorescence and flow cytometric analysis techniques wereused to observe cell *** effects of adenosine receptor(A1,A2a,A3and nonspecific receptor)antagonists(8-cpt,DMPX,MRS 1191,and theophylline)and an adenosine transporter protein inhibitor(dipyridamole)on adenosine-in-duced cell apoptosis were *** membrane potential was ana-lyzed using DePsipher fluorescent staining,and caspase activity was detectedusing a Fluorometric assay kit and a fluorescence microplate ***:Adenosine significantly reduced cell viability in a dose-and *** cytotoxicity of adenosine was related to the induction of cell *** adenosine receptor antagonists had no effect on cell ***,dipyridamole significantly reduced the percentage of adenosine-induced apoptoticcells from 27.3% to 7.1%(P3 mmol/L adenosineincreased caspase-3 activity 3.5-fold;dipyridamole markedly decreased caspase-3 activity 1.6-fold,and decreased apoptotic cell *** HepG2 cells weretreated with 3 mmol/L adenosine,mitochondrial membrane potential and theactivity of caspase-8 or-9 remained ***:Our results suggestthat adenosine-induced apoptosis in HepG2 cells is related to intracellular eventsrsther than cell surface receptors,and that a caspase-3 cascade activation isrequired,which is not mediated via a mitochondrial pathway.

关键词： adenosins adenosine receptor apoptosis caspase HepG2 mitochondria

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Effect of bilateral testicular resection on thymocyte and its microenvironment in aged mice

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Asian Journal of Andrology 2001年第4期3卷 271-275页

作者： Xi-Yun WEI Jin-Kun ZHANG Jun LI Su-Biao CHEN Department of Anatomy,Histology and Embryology,shantou university medical college,shantou 515031,china Department of Anatomy Histology and Embryology Shantou University Medical College China. junkunzh@***

Aim: To observe the changes in thymocyte and its microenvironment in aged mice after bilateral testicular ***: In male old mice, at the 25th day after testicular resection, the peripheral blood and thymus were collect-ed . Blood and thymus suspension smears were prepared for quantitative histochemistry and immunohistochemistry studyunder light and electron microscopes. Results; In testes resected mice the size and the weight of thymus weremarkedly increased. The demarcation between cortex and medulla was clear. The cortex was thickened and the celldensity was increased. The ratio of cortex/medulla stereometry was increased. The total cell count, thymocyte count,the percentage of acid a-naphthyl acetate esterase (ANAE) positive thymocytes, nonlymphocytes and the rosette forma-tion of macrophages and thymocytes were all increased. The thymocytes surrounded closely to the light thymic epithelialcells, dendritic cells or macrophages. The lymphocytes, particularly the ANAE positive lymphocytes of peripheralblood were increased. Conclusion; After bilateral testicular resection, the thymus of aged male mice showed mor-phological regeneration and the thymocytes and its microenvironment appeared to be definitely improved. It is suggestedthat testicular resection may improve immune function. (Asian J Androl 2001 Dec; 3; 271 — 275)

关键词： testis orchiectomy thymocyte microenvironment mice

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Telomere and telomerase in the initial stage of immortalization of esophageal epithelial cell

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World Journal of Gastroenterology 2002年第2期8卷 357-362页

作者： Zhong-Ying Shen Li-Yan Xu Wei-Jia Cai Min-Hua Chen Jian Shen,Department of Tumor Pathology,medical college of shantou university,shantou 515031,Guangdong Province,china En-Min Li,Department of Biochemistry and Molecular Biology,medical college of shantou university,shantou 515031,Guangdong Province,china Yi Zeng,Institute of Virology,Chinese Academy of Preventive Medicine,Beijing 100052,china Department of Tumor Pathology Medical College of Shantou University 22 Xinling Road Shantou 515031 Guandong Province China. zhongyingshen@***

AIM: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization. METHODS: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E(6)E(7) genes of human papillomavirus (HPV) type 18 in our laboratory. Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TRAP). Expressions of subunits of telomerase, hTR and hTERT, were assessed by RT-PCR. DNA content in cell cycle was detected by flow cytometry. The cell apoptosis was examined by electron microscopy (EM) and TUNEL label. RESULTS: SHEE cells from the 6th to 10th passages showed cellular proliferation with a good differentiation. From the 12th to the 16th passages, many senescent and apoptotic cells appeared, and the telomere length sharply shortened from 23kb to 17kb without expression of hTERT and telomerase activity. At the 20th passage, SHEE cells overcame the senescence and apoptosis and restored their proliferative activity with expression of telomerase and hTERT at low levels, but the telomere length shortened continuously to the lowest of 3kb. After the 30th passage cells proliferation was restored by increment of cells at S and G2M phase in the cell cycle and telomerase activity expressed at high levels and with maintenance of telomere length. CONCLUSION: At the early stage of SHEE cells, telomeres are shortened without expression of telomerase and hTERT causing cellular senescence and cell death. From the 20th to the 30th passages, the activation of telomerase and maintenance of telomere length show a progressive process for immortalization of esophageal epithelial cells. The expression of telomerase may constitute a biomark

关键词： cell Transformation, Neoplastic Apoptosis cell Division cell Line cell Size Epithelial cells Esophagus Humans In Situ Nick-End Labeling Papillomavirus, Human Research Support, Non-U.S. Gov't Telomerase Telomere

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