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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 Jinan 250012, China 2 Department of Medicine, Medical Molecular Biology Experimental Center, shandong University Jinan 250012 China Department of Thoracic surgery Qi Lu HospitalShandong UniversityJinan250012China Department of Medicine Medical Molecular Biology Experimental CenterShandongUniversityJinan 250012China

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

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Polyamine Accumulation in Transgenic Tomato Enhances the Tolerance to High Temperature stress

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Journal of Integrative Plant Biology 2009年第5期51卷 489-499页

作者： Lin Cheng Yijing Zou shuli Ding Jiajing Zhang Xiaolin Yu Jiashu Cao Gang Lu Key Laboratory of Horticultural Plant Growth Development and Biotechnology Ministry of Agriculture Department of Horticulture Zhejiang University Hangzhou 310029 China

Polyamines play an important role in plant response to abiotic stress. s-adenosyl-l-methionine decarboxylase （sAMDC） is one of the key regulatory enzymes in the biosynthesis of polyamines. In order to better understand the effect of regulation of polyamine biosynthesis on the tolerance of high-temperature stress in tomato, sAMDC cDNA isolated from saccharomyces cerevisiae was introduced into tomato genome by means of Agrobacterium tumefaciens through leaf disc transformation. Transgene and expression was confirmed by southern and Northern blot analyses, respectively. Transgenic plants expressing yeast sAMDC produced 1.7- to 2.4-fold higher levels of spermidine and spermine than wildtype plants under high temperature stress, and enhanced antioxidant enzyme activity and the protection of membrane lipid peroxidation was also observed. This subsequently improved the efficiency of CO2 assimilation and protected the plants from high temperature stress, which indicated that the transgenic tomato presented an enhanced tolerance to high temperature stress （38℃） compared with wild-type plants. Our results demonstrated clearly that increasing polyamine biosynthesis in plants may be a means of creating high temperature-tolerant germplasm.

关键词： heat stress Lycopersicon esculentum polyamines saccharomyces cerevisiae s-adenosylmethionine decarboxylase transformation.

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Prostate specific Antigen Promoter-Driven Adenovirus-Mediated Expression of Both ODC and AdoMetDC Antisenses Inhibit Prostate Cancer Growth

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Chinese Journal of Cancer Research 2010年第3期22卷 224-230页

作者： Wei Li Hui Xiong Yi-lin Hong Chun-hua Zhang Chang-chun Liu Biomedical Engineering Institution School of Control Science and EngineeringShandong University Jinan 250061 China Minimally lnvasive Urology Center Shandong Provincial Hospital Affiliated toShandong University Jinan 250021 China

Objective：To generate recombinant adenovirus that could simultaneously express ornithine decarboxylase（ODC） and s-adenosylmethionine decarboxylase（AdoMetDC） antisenses specifically in prostate cancer cells,and evaluate its inhibitory effect on prostate cancer in ***：Fragments of ODC and AdoMetDC genes were generated by PCR,cloned into the pPGL-PsEs,and then recombined with pAdEasy-1 vectors in AdEasy-1 ***-PsEs-ODC-AdoMetDCas virus was produced in HEK293 *** transfection with Ad-PsEs-ODC-AdoMetDCas,the levels of ODC or AdoMetDC were determined by RT-PCR and western blot *** effect of Ad-PsEs-ODC-AdoMetDCas treatment on tumor formation and growth was evaluated in xenograft models of prostate cancers in ***：The plasmid pAdEasy-PsEs-ODC-AdoMetDCas was successfully constructed and the recombinant Ad-PsEs-ODC-AdoMetDCas adenovirus was *** with Ad-PsEs-ODC-AdoMetDCas adenovirus significantly inhibited the expression of ODC and AdoMetDC genes specifically in prostate DU145 cells,but not H1299,HT29 and HepG2 cancer cells,and disrupted the ability of DU145 cells to form solid prostate cancer in *** treatment with Ad-PsEs-ODC-AdoMetDCas adenovirus significantly inhibited the growth of engrafted prostate tumors in ***：The recombinant Ad-PsEs-ODC-AdoMetDCas adenovirus specifically reduces the expression of both ODC and AdoMetDC genes in prostate cells and may be used for treatment of prostate cancers at the clinic.

关键词： Prostate androgen independent promoter Ornithine decarboxylase s-adenosylmethionine decarboxylase Adenovirus Prostate cancer

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Expression of sAMDC Gene for Enhancing the shelf Life for Improvement of Fruit Quality Using Biotechnological Approaches into Litchi (Litchi chinensis sonn.) Cultivars

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Advances in Bioscience and Biotechnology 2016年第6期7卷 300-310页

作者： Dilip Kumar Das Manoj Prabhakar Dipti Kumari Nutan Kumari Post Graduate Department of Biotechnology T.M. Bhagalpur University Bihar India

Polyamines play an important role in plant response to abiotic stress. s-adenosyl-1-methionine decarboxylase (sAMDC) is one of the key regulatory enzymes in the biosynthesis of polyamines. In order to better understand the effect of regulation of polyamine biosynthesis on the shelf life improvement of litchi fruit, sAMDC cDNA isolated from Datura stramonium cloned in pBI121 was introduced into litchi genome by means of Agrobacterium tumefaciens through zygote disc transformation. Transgene and its expression are confirmed by southern and Northern blot analyses, respectively. Transgenic plants expressing Datura sAMDC produced 1.7- to 2.4-fold higher levels of spermidine and spermine than wildtype plants under normal environmental condition, which indicated that the transgenic litchi presented an enhanced polyamines synthesis compared to wildtype plants. Our results demonstrated clearly that increasing polyamine biosynthesis in plants may be a means of creating improved fruit shelf life germplasm.

关键词： shelf Life Litchi chinensis sonn. Datura stramonium s-adenosylmethionine decarboxylase Polyamines Transformation

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