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Construction of a regulable gene therapy vector targeting for hepatocellular carcinoma

World Journal of Gastroenterology 2003年第4期9卷 688-691页

作者： Shao-YingLu Yan-FangSui Zeng-ShanLi Cheng-EnPan JingYe Wen-YongWang DepartmentofGeneralSurgery FirstHospitalofXi'anJiaotongUniversityXi'an710061ShaanxiProvinceChina DepartmentofPathology FourthMilitaryMedicalUniversityXi'an710032ShaanxiProvinceChina DepartmentofGeneralSurgery FirstHospitalofXi'anJiaotongUniversityXi'an710061ShaanxiProvinceChina

AIM: To construct a gene modified hepatocellular carcinoma (HCC) specific EGFP expression vector regulated by abbreviated cis-acting element of AFP ***: The minimal essential DNA segments of AFP gene enhancer and promoter were synthesized through PCR from Genome DNA of HepG2 cells. Gene fragments were then cloned into the multiple cloning site of non-promoter EGFP vector pEGFP-t. Recombinant plasmid was transferred into positive or negative AFP cell lines by means of lipofectamine. The expression of EGFP was tested by fluorescence microscope and flow cytometry. The effect of all-trans retinoic acid (ATRA) on the expression of EGFP was tested in different ***: By the methods of restriction digestion and sequence analyses we confirmed that the length, position and orientation of inserted genes of cis-acting element of AFP were all correct. The transcription of EGFP was under the control of AFP cis-acting element. The expressing EGFP can only been detected in AFP producing hepatoma *** expression rate of EGFP in G418 screened cell line was 34.9±4.1%. 48 h after adding 1×10-7M retinoic acid, EGFP expression rate was 14.7±3.5%. The activity of AFP gene promoter was significantly suppressed by addition of 1×10-7M retinoic acid (P＜0.05, P=0.003, t=6.488).CONCLUSION: This recombinant expression vector can be used as a gene therapy vector for HCC. The expression of tumor killing gene will be confined within the site of tumor and the activity of which can be regulated by retinoic acid.

关键词：肝细胞癌可控基因疗法载体聚合酶链反应 DNA扩增

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Superantigen-SEA gene modified tumor vaccine for hepatocellular carcinoma:An in vitro study

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World Journal of Gastroenterology 2004年第1期10卷 53-57页

作者： Shao-YingLu Yan-FangSui Zeng-ShanLi Jing-Ye Hai-LongDong PingQu Xiu-MinZhang Yu-SongLi DepartmentofPathology FourthMilitaryMedicalUniversityXi'an710032ShaanxiProvinceChina DepartmentofPathology FourthMilitaryMedicalUniversityXi'an710032ShaanxiProvinceChina

AIM: To construct an eukaryotic superantigen gene expression vector containing the recombinant gene of SEA and CD80 molecule transmembrane region (CD80TM), and to expresss taphylococcus enterotoxin A (SEA) on the membrane of hepatocellular carcinoma (HCC) cell to form a superantigen gene modified tumor vaccine for HCC. METHODS: SEA and linker-CD80TM gene were amplified through PCR from plasmid containing cDNA of SEA and *** fragments were then subcloned into the multiple cloning sites of retroviral vector pLXSN. Recombinant plasmid was transferred into HepG2 cells mediated with lipofectamine,positive clones were selected in culture medium containing G418. RT-PCR and indirect immunofluorescence studies confirmed that SEA was expressed specifically on HCC cell membrane. INFT-ELISPOT study demonstrated that SEA protein was expressed on the membrane of HCC *** of HepG2-SEA primed CTLs (SEA-T) was analyzed by^51Cr release assay. T cells cultured with rhIL-2 (IL-2-T) were used as control. RESULTS: Restriction digestion and sequence analyses confirmed the correctness of length, position and orientation of inserted fusion genes. SEA was expressed on the surface of HepG2 cells, HepG2-SEA had strong stimulating effect on production of HepG2 specific CTL (P<0.001). SEA-T had enhanced cytotoxicity to HepG2 cells (P<0.05). CONCLUSION: Tumor cell membrane expressed superarfdgen can be used to reinforce the immune effect of tumor cell vaccine for HCC, which provides a new method of the enhanced active immunotherapy for HCC.

关键词：超抗原-SEA基因肿瘤疫苗肝细胞癌体内试验免疫疗法

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肝癌超抗原/共刺激分子免疫基因治疗研究

肝癌超抗原/共刺激分子免疫基因治疗研究

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中国科协第五届青年学术年会

作者：李增山隋延仿中国人民解放军第四军医大学病理教研室中国人民解放军第四军医大学病理教研室

人们已经认识到肿瘤细胞存在抗原性弱、缺乏第二信号(共刺激分子)作用等问题,无法激活机体有效的免疫排斥反应,从而导致其免疫治疗效果不理想。已有的研究表明给肿瘤细胞转染免疫刺激分子编码基因,使其表达免疫刺激分子,

关键词：肝癌超抗原葡萄球菌肠毒素A CD80 免疫基因治疗

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