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Primary targeting of recombinant fv-immunotoxin hscfv_(25)-mTNFα against hepatocellular carcinoma

World Journal of Gastroenterology 2004年第13期10卷 1872-1875页

作者： JingZhang Yan-FangLiu Shou-JingYang QingQiao HongCheng Chuan-ShanZhang Fu-ChengMa Hua-ZhangGuo DepartmentofPathology XijingHospitalFourthMilitaryMedicalUniversityXi'an710033ShaanxiProvinceChina DepartmentofGeneralSurgery TangduHospitalFourthMilitaryMedicalUniversityXi'an710038ShaanxiProvinceChina

AIM: To obtain human recombinant fv-immunotoxin hscfv25-mTNFα (mutant human TNFαfused to human scfv25) against hepatocellular carcinoma (HCC).METHODS: Two relevant sites of enzymatic digestion were added to mTNFα by PCR. mTNFα was linked to the 3' end of hscfv25 in pGEX4T-1 vector. This anti-HCC recombinant fv-immunotoxin hscfv25-mTNFα was expressed in Escherichia coliand purified from inclusions. After purified by glutathione-S-transferase affinity chromatography and thrombin digestion, it was identified by electrophoresis and Western blot. And then, the purified recombinant fvimmunotoxin was injected into nude mice with HCC xenografts through their tail veins, mTNFα protein and PBS were used as control at the same time. After treated for two weeks, nude mice were executed. The bulk and weight of tumors were observed. The tumor tissues were stained by immunohistochemical method with TNFα ***: The expression ratio of recombinant fv-immunotoxin hscfv25-mTNFα was 12% of bacterial protein. The result of tumor restraining trials of hscfv25-mTNFα showed 2/5 complete remission and 3/5 partial remission, mTNFα restraining trials showed 5/5 partial remission. The therapeutic result of hscfv25-mTNFα was better than that of mTNFα (F=8.70, Pfv25-mTNFα remedial tumor tissues were positive for TNFα by immunohistochemical staining. The positive granules mainly existed in the cytoplasm of tumor ***: Recombinant fv-immunotoxin hscfv25-mTNFα has better therapeutic effect than mTNFα. It can inhibit the cellular growth of HCC and has some potential of clinical application.

关键词：重组器官 fv-抗毒素 hscfv25-mTNFα 肝细胞癌肿瘤 HCC

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