Construction and Study of Specific HSVTK/GCV Reconstructed AdenoAssociated Virus Plasmid of Human Hepatocellular Carcinoma_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LI Zhihui , YAN Lnan , ZHAO Yongheng , LIU Zhimin , LIU Ziming , LI Yu.

Department of General Surgery， West China Hospital， Sichuan University， Chengdu 610041, China;

Corresponding author：

Keywords：

Target gene therapyAdenoassociated virusHepatocellular carcinoma

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ObjectiveTo investigate target gene therapy for hepatocellular carcinoma (HCC). MethodsHerpes simplex virus thymidine kinase (HSVTK) gene was inserted into the gene of AFP enhancer/ALB promoter with adenoassociated virus (AAV) plasmid (WAV2) as a carrier, and a hybrid plasmid pWAV2/AFPALB/HYTK was constructed. Besides, plasmid pEGFP1/AFPALB was also constructed. Two kinds of plasmids were transferred into AFP positive cells HepG2 and AFP negative cells 7721, SPC and 7901.ResultsIt was found that enhance green fluorescence protein could only be seen in AFP positive cells HepG2. 710 bp DNA was amplified only in AFP positive HepG2 cells.ConclusionPlasmid pWAV2/AFPALB/HYTK for HCC demonstrates specificity in vitro.

Citation： LI Zhihui,YAN Lnan,ZHAO Yongheng,LIU Zhimin,LIU Ziming,LI Yu.. Construction and Study of Specific HSVTK/GCV Reconstructed AdenoAssociated Virus Plasmid of Human Hepatocellular Carcinoma. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2004, 11(3): 234-235. doi: Copy

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1. Hermonat PL, Muzyczka N. Use of adenoassociate virus as a mammalian DNA cloning vector: transduction of neomycin resistance into mammalian tissue culture cells [Ｊ]. Proc Natl Acad Sci USA, 1984; 81(20)∶ 6466.
2. 伍志坚，吴小兵，侯云德. 系列腺病毒伴随病毒载体的构建及表达β半乳糖苷酶的研究 [Ｊ]. 病毒学报， 2000; 16(1)∶1.
3. Bilbao R, Gerolami R, Bralet MP, et al. Transduction efficacy, antitumoral effect, and toxicity of adenovirusmediated herpes simplex virus thymidine kinase/ganciclovir therapy of hepatocellular carcinoma: the woodchuck animal model [Ｊ]. Cancer Gene Ther, 2000; 7(5)∶657.
4. Kaneko S, Hallenbeck P, Kotani T, et al. Adonovirusmediated gene therapy of hepatocellular carcinoma using cancer specific gene expression [Ｊ]. Cancer Res, 1995; 55(22)∶5283.
5. Watanabe K, Saito A, Tamaoki T. Cellspecific enhancer activity in a far upstream region of the human alphafetoprotein gene [Ｊ]. J Biol Chem, 1987; 262(10)∶4812.
6. Fumihiko K, Yasushi S, Yoko Y, et al. Gene therapy for αfetoproteinproducing human hepatoma cell by adenovirusmediate transfer of the herpes simplex virus thymidine kinasegene [Ｊ]. Hepatology, 1996; 23(6)∶1359.
7. Hanke P, Serwe M, Dombrowski F, et al. DNA vaccination with AFPencoding plasmid DNA prevents growth of subcutaneous AFPexpressing tumors and does not interfere with liver regeneration in mice [Ｊ]. Cancer Gene Ther, 2002; 9(4)∶346.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction and Study of Specific HSVTK/GCV Reconstructed AdenoAssociated Virus Plasmid of Human Hepatocellular Carcinoma

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