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Co-application of ricin A chain and a recombinant adenovirus express-ing ricin B chain as a novel approach for cancer therapy

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Acta Pharmacologica Sinica 2007年第5期28卷 657-662页

作者： Hong-bin WANG~(1,3),Fei XIA~(1,3),Jing GE~1,Juan YIN~1,Li-song TAN~4,Pei-de ZHANG~2,Jiang ZHONG~(1,5)Departments of ~1Mierobiology and Microbial Engineering and ~2Biochemistry and Molecular Biology,School of Life Sciences,FudanUniversity,Shanghai 200433,China ~3Marine College,Huaihai Institute of Technology,Lianyungang 222005,China ~4Shanghai PulmonaryHospital,Shanghai 200433,China Department of Microbiology and Microbial Engineering School of Life Sciences Fudan University Shanghai China Department of Marine College Huaihai Institute of Technology Lianyungang China Department of Shanghai Pulmonary Hospital Shanghai China Department of Biochemistry and Molecular Biology School of Life Sciences Fudan University Shanghai China

Aim:To develop a novel ricin-based approach for the safe and effective therapyof ***:The ricin A chain （RTA） was expressed in Escherichia coli inthe form of a 6×His-tagged fusion protein and purified with Ni2+-NTA affinityresin.A replication-deficient ricin B chain （RTB）-expression adenovirus greenfluorescence protein （AdGFP-RTB） was *** and AdGFP-RTB weretested for cytotoxicity either individually or in combination in human cell linesHEK293,HeLa,SMMC7721,and *** viability was determined with trypanblue staining or MTT ***:The expression and release of RTB,as wellas the entry of RTA into AdGFP-RTB-infected cells were *** RTAand AdGFP-RTB was used individually,neither was toxic to the *** theywere applied together,significant cell death was observed in all of the cell *** cell-killing effect correlated with the amount of RTA protein used,withcell mortality at about 60% at 4.8 μg RTA in combination with AdGFP-RTB at 100pfu/*** major cell killing was seen when RTA was used in combination with acontrol adenovirus *** treatment of healthy HeLa cells with the virus-free supernatant from AdGFP-RTB/RTA-treated HeLa cells resulted in cell death,suggesting the formation of RTA/RTB complex,and a potential by-stander ***:The new approach was successful in *** modifications ofthe adenovirus vector,as well as an in vivo study are needed to confirm its poten-tial in cancer therapy.

关键词： ricin ribosome inactivating protein protein expression recombinant adenovirus cell killing cancer therapy

Effect of Interstitial Chemotherapy with ricin Temperature-Responsive Gel for Anti-Breast Cancer and Immune Regulation in Rats

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Chinese Journal of Integrative Medicine 2013年第1期19卷 48-53页

作者：陈志奎林礼务蔡晶杨发端蔡华晶薛恩生黄静魏洪芬张秀娟 Ultrasonic Department Affiliated Union Hospital of Fujian Medical University Department of Pathology Affiliated Xiehe Hospital of Fujian Medical University Integrative Medical Institute of Fujian University of Traditional Chinese Medicine Department of Pharmacology College of PharmacyFujian Medical University

Objective： To explore the effect of ricin temperature response gel on breast cancer and its regulatory effect on immune function in rats. Methods： ricin was purified by chromatography and identified by immunoblotting. The rat subcutaneously transplanted breast cancer model was established. Forty model rats with a tumor diameter of about 3.0 cm were subjected to the study. They were randomized into four groups equally： the model group and three treated groups （blank gel, ricin, ricin-gel） were administered with blank gel, ricin, and ricin temperature response gel via percutaneous intratumor injection, respectively. The tumor was isolated 10 days later for the estimation of tumor inhibition rate （FIR） by weighing, pathologic examination, and detection of tumor apoptosis-associated genes bcl-2 and bax with semiquantitative RT-PCR. Also, peripheral blood was obtained to test T-lymphocyte subsets, the killing function of lymphocytes, and the contents of tumor necrosis factor- oL （TNF- oL ） and interleukin-2 （IL-2）. The outcomes were compared between groups. Results： The TIR in the ricin-gel group was 61.8%, with the pathologic examination showing extensive tumor tissue necrosis. Compared with the model group, after ricin temperature response gel treatment, bcl-2 expression was down- regulated, bax expression was up-regulated, CD4＋ lymphocytes and CD4＋/CD8＋ ratio in peripheral blood were increased, the killing function of lymphocytes was enhanced, and the contents of TNF- α and IL-2 were elevated （P〈0.05 or P〈0.01）. Conclusion： Intratumor injection of ricin temperature-responsive gel showed significant antitumor effect on breast cancer and could enhance the immune function in the tumor-bearing rat.

关键词： ricin gel breast cancer mesenchymal chemotherapy immune regulation

Interstitial chemotherapy with ricin-loaded thermosensitive hydrogel in pancreatic cancer xenograft

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Hepatobiliary & Pancreatic Diseases International 2009年第4期8卷 418-423页

作者： Chen, Zhi-Kui Lin, Li-Wu Weng, Xiu-Hua Xue, En-Sheng Lin, Yong-Hua Fujian Med Univ Union Hosp Dept Ultrasound Fuzhou 350001 Peoples R China Fujian Med Univ Affiliated Hosp 1 Dept Pharm Fuzhou 350001 Peoples R China Fuzhou Gen Hosp Dept Gen Surg Fuzhou 350025 Peoples R China

BACKGROUND: Pancreatic cancer is one of the most aggressive malignancies, and has a poor prognosis. Despite efforts made in multiple fields, there has been little success in improving the disease-free survival rate of patients. This study was undertaken to investigate the effectiveness and feasibility of using intra-tumoral injection of ricin-loaded thermosensitive hydrogel for treatment of pancreatic cancer xenografts, attempting to develop a new treatment for human pancreatic cancer. METHODS: BALB/c-(nu/nu) nude mice were inoculated subcutaneously in the right flank with the human pancreatic cancer cells, SW1990. Fourteen days after inoculation, 32 mice, bearing tumors of volume 1.5-2.0 cm(3), were randomly assigned to one of four groups, and given an intra-tumoral injection of: (1) saline; (2) 23% w/w thermosensitive hydrogel alone; (3) ricin, 10 mu g/kg; or (4) 10 mu g/kg ricin loaded in thermosensitive hydrogel. On day 14 after administration, the tumors were excised to calculate the inhibition rate of tumor growth and perform histopathological examination. Tumor cell apoptosis was detected by flow cytometry, and RT-PCR was performed to evaluate the mRNA expression levels of Bc12 and Bax. RESULTS: Intra-tumoral injection of ricin-loaded thermosensitive hydrogel resulted in remarkable control of tumor growth. The tumor became necrotic by day 14 after administration. The histological results clearly confirmed that the tumor cells were lysed. The percentage of apoptotic cells detected by flow cytometry was higher in the ricin hydrogel group than in the other groups. Semi-quantitative RT-PCR revealed that the mRNA expression level of Bc12 was down-regulated whereas Bax was upregulated. CONCLUSIONS: Intra-tumoral injection of ricin-loaded thermosensitive hydrogel may provide an effective approach for interstitial chemotherapy in pancreatic cancer. Inducing apoptosis by downregulating Bcl2 expression and upregulating Bax expression may be a key molecular mechanism.

关键词： pancreatic neoplasm ricin hydrogel therapeutics drug delivery system

Modification of ricin and its hepatotoxicity and activity against hepatocellular cancer in mice

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World Journal of Gastroenterology 1998年第4期4卷 35-38页

作者： WANG WenXue, DONG JuYing, ZHOU SiYuan, LI WenLi and ZHAO YingKeywords ricin/chemistry ricin/toxicity liver/drug effects liver neoplasms carcinoma, hepatocellular glutathione transferase

AIM To observe the effects of ricin (RT) with and without chemical modification on both hepatotoxicity of mice and activity against hepatocellular cancer (HCC), and evaluate the possibility to improve RT anticancer activity via chemical *** RT was modified with N-succinimidyl3-(2-pyridyldithio) propionate (SPDP), a heterobifunctional cross-linker, and SPDP derivative of RT (PDP-R) was obtained. The serum glutathione-s-transferase (SGST) activity, as an index of liver damage, was determined in mice intoxicated with RT and PDP-R, at various doses and time. The tissue damage of HCC in the nude mice ip injected with PDP-R was compared with that with RT at the same dose by immunohistochemical method, the relative content of both RT and PDP-R in the HCC tissues was measured by computerized *** The SGST activities increased with doses or/and time intoxicated with both RT and PDP-R, and the increase in the value of RT group was more significant than that in the PDP-R group; the SGST activity of RT group was 2.8-fold (P＜0.01) of PDP-R group at a dose of 12.5μg/kg for 42h, showing the much lower toxicity of R-PDP than that of RT. Under an optical microscope, hemolysis and necrosis of massive cells in the HCC tissues of PDP-R group were observed and the ratio of necrosis mounted to 90.5% while the corresponding value of RT group only to 62.5%. With computerized image-analysis, the average relative content of RT and PDP-R in the HCC tissues, represented as greyness, was 140.06±3.43 and 169.10±2.74, respectively. There was significant difference between the two (P＜0.05), indicating the higher content of PDP-R in the HCC tissue than that of *** The hepatotoxicity of PDP-R to mice may be reduced by chemical modification with SPDP, but both the affinity of PDP-R to the HCC tissues and ability to kill it may be stronger than that of RT. So this might be a valuable attempt to improve the anticancer activity of RT.

关键词： ricin Modification

Development of colloidal gold-based immunochromatographic assay for the rapid detection of ricin toxin

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Development of colloidal gold-based immunochromatographic as...

Protective effects of anti-ricin A-chain antibodies delivered intracellularly against ricin-induced cytotoxicity

第九届中国生物毒素学术研讨会

作者： Jing-Hong WANG,Lin KANG,Shan GAO,Chong NIE,Jing-Lin WANG Laboratory of Toxinology,The State Key laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Beijing 100071, China

＜正＞A rapid,signal step and highly sensitive immunochromatographic（IC） assay was developed to detect *** assay was based on the sandwich format using monoclonal antibody（Mab） and polyclonal antibody（Pab） of two distinct *** anti-ricin A chain Mab was immobilized as a capture antibody on a porous nitrocellulose membrane where goat anti-rabbit IgG（control line） was separately coated with,while the other anti-ricin B chain Pab was conjugated to colloidal gold particles that served as a detector *** ricin-containing sample was added to the membrane and allowed to react with Pab-coated *** mixture was then passed along the porous membrane by capillary action past the Mab in the detection zone,which will bind the particles that had ricin bound to their surfaces,giving a red color within this detection zone with an intensity proportional to ricin *** the absence of ricin,no immunogold was bound to the solid-phase antibody. With this method,50ng/mL of ricin was detected in less than 10 min,and the assay also showed no cross reaction to ricin toxin.

关键词： Colloidal gold Immunochromatographic assay ricin Antibody

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World Journal of Biological Chemistry 2010年第5期1卷 188-195页

作者： Frank Martiniuk Seth Pincus Sybille Müller Heinz Kohler Kam-Meng Tchou-Wong Department of Medicine New York University School of Medicine550 First AvenueNew YorkNY 10016 United States Department of Research Children's Hospital200 Henry Clay AvenueNew OrleansLA 70118United States Immpheron Inc.5235 Athens-Boonesboro RoadLexingtonKY 40509United States Markey Cancer Center University of Kentucky800 Rose StreetLexingtonKY 40536-0096United States Department of Environmental Medicine New York University School of Medicine57 Old Forge RoadTuxedoNY 10987United States

AIM:To evaluate the ability of anti-ricin A-chain antibodies,delivered intracellularly,to protect against ricininduced cytotoxicity in RAW264.7 cells. METHODS:Anti-deglycosylated ricin A-chain antibody and RAC18 anti-ricin A-chain monoclonal antibody were delivered intracellularly by encapsulating in liposomes or via conjugation with the cell-penetrating MTS-transport ***264.7 cells were incubatedwith these antibodies either before or after ricin *** changes in cytotoxicity were estimated by MTT ***-localization of internalized antibody and ricin was evaluated by fluorescence microscopy. RESULTS:Internalized antibodies significantly increased cell viability either before or after ricin exposure compared to the unconjugated *** microscopy confirmed the co-localization of internalized antibodies and ricin inside the cells. CONCLUSION:Intracellular delivery of antibodies to neutralize the ricin toxin after cellular uptake supports the potential use of cell-permeable antibodies for postexposure treatment of ricin intoxication.

关键词： ricin inhibitors Antibody internalization Cell-penetrating peptide Cell-permeable antibodies Liposome-encapsulated antibodies

ricin引起外周神经跨越神经节溃变的超微结构研究

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神经解剖学杂志 1992年第1期8卷 129-133页

作者：杜彦军秦秉志李继硕第四军医大学解剖学教研室西安710032

为了探索神经毒剂ricin对神经元溃变过程中的效用及最佳用量的效果,将10%ricin注射于坐骨神经分支中,按梯次剂量将实验动物分组,分别于术后不同存活时间观察后根节神经元的溃变状态及过程,并在电镜下观察骶髓后连合核内的溃变终末像。... 详细信息

为了探索神经毒剂ricin对神经元溃变过程中的效用及最佳用量的效果,将10%ricin注射于坐骨神经分支中,按梯次剂量将实验动物分组,分别于术后不同存活时间观察后根节神经元的溃变状态及过程,并在电镜下观察骶髓后连合核内的溃变终末像。结果表明注射6μl的10%ricin、存活5-7天的材料,初级传入终末溃变的超微结构像最佳。ricin所致的跨节溃变终末呈现多型性变化。

关键词： ricin 超微结构跨越神经节溃变

Structural insights into the interaction of the ribosomal P stalk protein with a type II ribosome-inactivating protein ricin

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Structural insights into the interaction of the ribosomal P ...

Screening and comparison of antitoxic activity of five novel monoclonal antibodies against ricin poisoning

中国生物化学与分子生物学会2016年全国学术会议

作者： Xiaojiao Fan Yuwei Zhu Chongyuan Wang Liwen Niu Maikun Teng Xu Li Hefei National Laboratory for Physical Sciences at Microscale Innovation Center for Cell Signaling Network School of Life ScienceUniversity of Science and Technology of China Key Laboratory of Structural Biology Hefei Science Center of CAS Chinese Academy of cience

ricin is a type II ribosome-inactivating protein(RIP).It depurinates the A4324 at the sarcin-ricin loop of 28S r RNA,which thereby inactivates ribosome by preventing elongation factors binding to the GTPase activation *** studies have disclosed that the conserved C-terminal domain(CTD)of eukaryotic ribosomal P stalk proteins is involved in the targeting

关键词： ribosome-inactivation protein ricin ribosomal P stalk protein sarcin-ricin loop

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Screening and comparison of antitoxic activity of five novel...