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Cloning of chicken anemia virus vp3 Gene and Apoptosis Inductive Effect of vp3 Gene In Vitro

Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第4期23卷 329-331,334页

作者：孙军王宇哲宗义强屈伸 Department of Biochemistry and Molecular Biology

Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE TM mediated transfection in vitro with pcDNA vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L 02, and RT PCR was performed afterward. The results of RT PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.

关键词： chicken anemia virus gene cloning apoptosis vp3 gene

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Experimental Study on the Antitumor Effect of chicken anemia virus vp3 Gene against Liver Carcinoma In Vivo

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第2期23卷 105-107,115页

作者：申志发王宇哲宗义强屈伸 Department Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Tongji Medical CollegeHuazhong University of Science and Technology Wuhan 430030 Department of Biochemistry and Molecular Biology School of Basic Medical SciencesTongji Medical CollegeHuazhong University of Science and Technology Wuhan 430030

In order to testify the antitumor effect, especially its effect against liver carcinoma in vivo, of VP3 protein, one kind of protein coded by chicken anemia virus, recombinants pcDNA-vp3 containing chicken anemia virus vp3 gene, and control vector pcDNA3 were mixed with murine liver carcinoma cell lines H22 respectively. The mixture was injected subcutaneously into Balb/C mice. Some days later, the mice were killed and the solid tumor weighed. The antitumor efficiency was evaluated. The manners of VP3 protein in vivo inducing tumor cell death were identified by using TUNEL assay. All the results suggested that the injection of pcDNA-vp3 and H22 mixture resulted in a significant reduction of tumor growth in mice when compared with the results of control groups. TUNEL assay revealed that VP3 induced apoptosis in vivo. All these indicated that CAV vp3 might be a potential new gene in reducing the growth rate of tumor cells in liver carcinoma or in other kind of solid tumors in vivo.

关键词： chicken anemia virus VP3 protein apoptosis gene therapy

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Anti-tumor Effects of a Recombinant Fowlpox virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro

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Chemical Research in Chinese Universities 2011年第4期27卷 646-650页

作者： ZHU Ji-hong LI Xiao SUN Li-li ZHANG Mu-chun KAN Shi-fu LIU Lei HUANG Hai-yan YANG Guo-hua PIAO Bing-guo JIN Ning-yi Department of Gynecology and Obstetrics the First Hospital of Jilin University Changchun 130021 P. R. China Laboratory of Genetic Engineering of PLA Academy of Military Medical Sciences Changchun 130062 P. R. China Department of Head and Neck Surgery Tumor Hospital of Jilin Province Changchun 130012 P R. China Department of Urology China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China Department of Gastroenterology the First Hospital of Jilin University Changchun 130021 P. R. China

Apoptin is a chicken anemia virus-derived,p53-independent,bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells,but not of normal diploid *** explore the application of apoptin in tumor gene therapy,we used a recombinant fowlpox virus expressing apoptin protein （vFV-Apoptin） to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma（HeLa） cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide（MTT） assay,acridine orage/ethidium bromide（AO/EB） and annexin V staining test,*** results show that vFV-Apoptin inhibites the proliferation of HeLa cells in vitro through inducing the apoptosis of HeLa cells,and the inhibition effect of vFV-Apoptin has a dose-effect and time-effect *** results of animal models show that vFV-Apoptin significantly inhibits tumor growth,extends the lifespan of animals and improves the mean *** results indicate that vFV-Apoptin has a potential application in the tumor gene therapy.

关键词： Apoptin gene chicken anemia virus Human cervical carcinoma Anti-tumor effect

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Circulation of Immunosuppressive viruses and Avian Encephalomyelitis virus in Backyard chicken Flocks

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Advances in Microbiology 2020年第5期10卷 203-213页

作者： Priscila de Castro Almeida Pricila Ribeiro Silva Borges Priscilla K.Koerich Roberta Torres de Melo Igor Alves Batista Eliane Pereira Mendonca Rogério Reis Silva Lillian Karla Silva Belchiolina Beatriz Fonseca Elanco Saúde Animal Sao PauloBrazil BRF S/A UberlândiaBrazil Faculdade de Medicina Veterinária da Universidade Federal de Uberlandia UberlandiaBrazil Faculdade de Medicina Veterinária da Universidade de Uberaba UberabaBrazil

The objective of this study was to evaluate the circulation of chicken anemia virus (CAV), Infectious Bursal Disease virus (IBDV), Avian Reovirus (ARV) and Avian Encephalomyelitis virus (AEV) in properties of backyard chickens and carry out an epidemiological analysis. We evaluated 200 samples of chickens from 19 backyard chicken property. Only one property (P10) did not present serological titers for the diseases evaluated. This property is close to industrial farms as well as the other properties, however, P10 remained a few years without the breeding of chicks and these were the first poultry to be housed on site. This reinforces the importance of the fallow period for poultry production. The prevalence of virus-seroreactive birds was 78% (156/200), 64.5% (129/200), 78% (156/200), 78% (156/200) for CAV, IBDV, ARV and, EA, respectively. All the free-range farms studied are within a radius of 500 meters to 6 Km away from some establishments of industrial poultry. There was a correlation between serological titers for CAV and the frequency of disease in poultry (r = 0.6178). In places where birds are frequently sick, 30.76% reported that the disease occurs in chicks, 30.76% in broilers, 23.07% in broiler chickens and 7.69% in birds of all ages. Birds get sick more often in the summer period. The owners reported that the most common signs of disease were respiratory signs (snoring and nasal discharge) (46.15%), diarrhea (30.76%), and paralysis of wings and/or paws (38.46%). There was a correlation between the presence of untreated water in the property and serological titers for ARV (r = 0.5576). This report draws attention not only to high serological prevalence for the viruses studied but also important epidemiological aspects of backyard chicken diseases that may indirectly influence the industrial production.

关键词： chicken anemia virus Infectious Bursal Disease virus Avian Reovirus Epidemiology

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