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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 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

The big bang of genome editing technology: development and application of the CRISPR/Cas9 system in disease animal models

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Zoological Research 2016年第4期37卷 191-204页

作者： Ming SHAO Tian-Rui XU Ce-Shi CHEN Faculty of Life Science and Technology Kunming University of Science and Technology Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province Kunming Institute of Zoology Chinese Academy of Sciences

Targeted genome editing technology has been widely used in biomedical studies. The CRISPR- associated RNA-guided endonuclease Cas9 has become a versatile genome editing tool. The CRISPR/Cas9 system is useful for studying gene function through efficient knock-out, knock-in or chromatin modification of the targeted gene loci in various cell types and organisms. It can be applied in a number of fields, such as genetic breeding, disease treatment and gene functional investigation In this review, we introduce the most recent developments and applications, the challenges, and future directions of Cas9 in generating disease animal model. Derived from the CRISPR adaptive immune system of bacteria, the development trend of Cas9 will inevitably fuel the vital applications from basic research to biotechnology and bio- medicine.

关键词： CRISPR/Cas9 Animal models genetherapy

GENETICALLY MODIFIED DENDRITIC CELLS INDUCED SPECIFIC CYTOTOXITY AGAINST HUMAN HCC CELLS IN VITRO

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Chinese Journal of Cancer Research 2004年第4期16卷 246-250页

作者：刘彬彬叶胜龙贺平郑宁赵燕孙瑞霞刘银坤汤钊猷 Liver Cancer Institute Zhongshan Hospital & Fudan UniversityShanghai 200032

Objective: to transduce the tumor associated antigen gene MAGE-1 and/or IL-12 gene into dendritic cells (DC) and to observe the in vitro cytotoxic effect induced by the genetically modified DC against the human hepatocellular carcinoma (HCC) cell line SMMC7721. Methods: the MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected at appropriate differentiation stage by LMSN and/or a recombinant adenovirus AdmiL-12, which containing murine IL-12 gene. The control groups included retrovirus LXSN transfected, adenovirus AdBGFP transfected and non-transfected DCs. The MAGE-1 gene expression was identified by western blot and the mIL-12 p70 secretion was detected by ELISA assay. The in vitro cytotoxicities against SMMC7721 induced by genetically modified and control groups of DC were tested by MTT assay. Results: The MAGE-1 expression was detected by a monoclonal antibody in DCs tranfected with LMSN but not in control groups. At 16 h, 24 h and 48 h after transfection with AdmIL-12, the concentration of the mIL-12 p70 in the culture medium was 580pg/106 cells, 960pg/106 cells and 1100pg/106 cells respectively. The mIL-12 p70 secretions were not detected in other groups. The lytic activity (as judged by % lysis) induced by each groups of DC was 94.2±5.2% (LMSN and AdmIL-12 cotransfected group), 78.9±3.6% (LMSN transfected groups), 52.6±9.7% (AdmIL-12 transfected group), 34.7±4.3% (LXSN transfected group), 36.3±3.8% (AdBGFP transfected group) and 3.9±2.0% (non-transfected group) respectively. Except for LXSN transfected and AdBGFP transfected group, the difference of the lytic activities between other groups were statistically significant (P<0.05). Conclusion: The MAGE-1 gene modified DCs can induce relatively specific cytotoxicty against SMMC7721 in vitro and thus suggested that those genetically engineered DCs have the potential to serve as novel vaccine for HCC. Transduction of exogenous IL-12 gene into DCs may further enhance DCs’

关键词： MAGE-1 IL-12 Dendritic cells Tumor vaccine genetherapy Hepatocellular carcinoma

基因转染间充质干细胞的临床治疗:特点与问题

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中国组织工程研究 2014年第14期18卷 2257-2262页

作者：李丽春李剑平辽宁省血液中心辽宁省沈阳市110044

背景:骨髓间充质干细胞因制备较易、具备有较强的自我增殖能力、稳定的生物学性状、低免疫原性、能在宿主脑中生存较长时间、易于转染外源性基因且有较高的转染率、对肿瘤细胞具有趋向性而成为一种较有希望的基因治疗的靶细胞。目的:利... 详细信息

背景:骨髓间充质干细胞因制备较易、具备有较强的自我增殖能力、稳定的生物学性状、低免疫原性、能在宿主脑中生存较长时间、易于转染外源性基因且有较高的转染率、对肿瘤细胞具有趋向性而成为一种较有希望的基因治疗的靶细胞。目的:利用不同基因转移的方法分析间充质干细胞用于临床多种难治性疾病基因治疗的优势和缺陷。方法:应用计算机检索2000年1月至2013年12月PubMed数据库、中国期刊全文数据库、万方数据库相关间充质干细胞的文章,中文检索词为"间充质干细胞,基因治疗",英文检索词为"mesenchymal stem cells,gene therapy"。共检索到文献2 114篇,最终纳入符合标准的42篇文献。结果与结论:基因治疗有多种,按基因操作分为:①基因修正和基因置换。②基因增强和基因失活。按靶细胞分为生殖细胞基因治疗和体细胞基因治疗。按给药途径分为活体外途径和活体内途径。相对与其他细胞骨髓间充质干细胞能在宿主脑中生存较长时间、易于转染外源性基因且有较高的转染率、对肿瘤细胞具有趋向性而成为一种较有希望的基因治疗的靶细胞。间充质干细胞作为基因治疗的载体其移植物抗宿主病、克隆氏病的治疗在国外已经进入到3期临床阶段。国内应用间充质干细胞治疗临床上一些难治性疾病也取得了明显的疗效。探索干细胞移植最佳治疗窗口,寻找适宜的基因剂量、移植细胞数量及合适的治疗时机,发挥最佳的治疗效果是有待解决的问题。

关键词：骨髓间充质干细胞临床治疗基因转染中国期刊全文数据库体细胞基因治疗移植物抗宿主病 genetherapy 外源性基因

A Combination Therapy of rAd-p53 with Chemotherapy for Advanced Oral Carcinoma by Selective Intraarterial Infusion

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A Combination Therapy of rAd-p53 with Chemotherapy for Advan...

Epstein-Barr virus-based vectors used in cancer gene therapy

第五次全国口腔颌面—头颈肿瘤学术研讨会

作者： Li Long-Jiang Wang Li-Juan Li Yi Han Bo Huang Ju Department of Oral and Maxillofacial Surgery, West China College of Stamotology, Sichuan University, Chengdu 610041, PR China West China Health Hospital, Sichuan University, Chengdu 610041, PR China

＜正＞Purpose: To evaluate the safety and therapeutic effects of selective intraarterial rAd-p53 infusion combined with chemotherapy for advanced oral carcinoma. Methods: 56 patients with advanced oral carcinoma underwent unilateral or bilateral retrograde selective catheterization of external carotid artery. rAd-p53 combined with chemotherapeutic agents was administrated to group I （20 patients）, rAd-p53 to group II （16 patients）, and chemotherapeutic agents to group III （20 patients）. The therapeutic response of primary tumor and metastatic lesion was evaluated by clinical exam and morphological measurement. Meanwhile, wt-p53, Bax and Bcl-2 were evaluated by immunohistochemistry before and after treatment. Results: The complete response rate （40%） of group I was higher than that （25% and 25%, respectively ） of group II and group III （p＜0.05）. The therapeutic effects were not found the significant difference between group II and III （p＞0.05）. 47 patients experienced some sort of side effects, which was mostly transient flu-like symptom, suppression of marrow or both. No replication-deficient virus was detected in patients’ serum, urine or sputum at any assay. The transactivation of Bax and inactivation of Bcl-2 were only observed in group I and II. Conclusion: Routine chemotherapy combined with rAd-p53 infusion can increase the curative effect and decrease the toxic implication of chemotherapy.

关键词： Oral carcinoma genetherapy Chemotherapy Intraarterial infusion.

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Epstein-Barr virus-based vectors used in cancer gene therapy

Co-expression of TRAIL and Smac delivered by an E1B-55K deleted oncolytic adenovirus containing a caspase-8 cleavage site (IETD) elicits complete elimination of hepatocellular carcinoma xenograft

2000全国肿瘤学术大会

作者：陈汉奎广州市肿瘤医院生物治疗室

＜正＞Epstein-Barr virus（EBV）is well known as a human tumor virusassociated with a wide variety of human malignancies,includingnasopharyngeal carcinoma （NPC）,a malignant epithelial tumorsignificantly prevalent in the southern China.EBV could notbe used simply as a vector in gene delivery by molecular scientists because of its large genome（170kb）and its potential to cause tu- mor.However,two components of EBV genome DNA are useful in the con-

关键词： Epstein-Barr virus Epstein-Barr virus-based vectors genetherapy

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Co-expression of TRAIL and Smac delivered by an E1B-55K dele...

Combination Adenovirus-Mediated HSV-tk/GCV and Antisense IGF-1 Gene Therapy for Rat Glioma

2012年浙江省血液病学年会

作者： Shibing Wang Yuan Tan Wen Lei Yigang Wang Xiumei Zhou Kangjian Zhang Liang Chu Xinyuan Liu Wenbin Qian Xinyuan Institute of Medicine and Biotechnology School of Life Science Zhejiang Sci-Tech University Institute of Hematology The First Affiliated Hospital College of Medicine Zhejiang University Laboratory of Molecular Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences

Combination target therapy is necessary for cancer treatment because tumors are genetically diverse. In our current study, we generated a novel E1B-55K deleted oncolytic adenovirus that harbors both TRAIL and Smac genes (ZD55-TRAIL-IETD-Smac). In this vector, a caspase-8 cleavage site (IETD) was introduced between the genes, which allows the production of TRAIL and Smac, respectively, within virusinfected cells after activation of caspase-8. In our system, treatment with ZD55-TRAIL-IETD-Smac produced equal expression of TRAIL and Smac, and resulted in significant cytotoxicity against various cancer cells with stronger efficacy than that of ZD55-TRAIL or ZD55-Smac alone. The expression of these two therapeutic genes in the infected cells was downregulated by a caspase-8 inhibitor (Z-IETD-FMK) and upregulated by 5-FU (a activator of caspase-8), suggesting that cleavage of TRAIL-IETD-Smac fusion protein by activation of caspase-8. In a s.c. xenograft hepatocellular carcinoma (HCC) model, the antitumor effect of ZD55-TRAIL-IETD-Smac was significantly stronger than that of ZD55-TRAIL and ZD55-Smac, respectively. Importantly, intratumoral injection of ZD55-TRAIL-IETD-Smac eliminated all xenograft tumors established from a human HCC cell line Bel-7404, resulting in long-term, tumor-free survival. In conclusion, our data suggest that oncolytic adenovirus incorporating two different therapeutic genes by introduction of the IETD sequence may be attractive candidates for clinical application in treatment of cancer.

关键词： TRAIL Smac co-expression IETD virotherapy genetherapy

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苏州医学院学报 2000年第7期20卷 597-601页

作者：李向东惠国桢卢大儒王琪徐露邱信芳薛京伦 Department of Neurosurgery The First Hospital Affiliated to Suzhou Medical College

Objective To investigate the effects of combination adenovirusmediated HSVtk/GCV system and antisense IGF1 gene therapy for rat glioma and analyze the mechanism.Methods Using the recombinant adenovirus vector,GCV killing effeciency after combined gene transfer of HSVtk and antisense IGF1 was observed in vitro.Rat glioma was treated with HSVtk/GCV and antisense IGF1 and the survival rate of rats was observed.Results C6 cells transfected with tk and antisense IGF1 gene were more sensitive to GCV than that transfected with tk gene alone.The survival of the combination gene therapy group was prolonged significantly and large amounts of CD+ 4,CD+ 8 lymphocytes were detected in the tumor tissues.Conclusion Antisense IGF1 gene may enhance the tumorkilling effects of HSVtk/GCV.

关键词： HSV-tk IGF-1 glioma genetherapy