人肝癌特異性HSVTK/GCV重組腺病毒相關病毒質粒的構建與研究_《中國普外基礎與臨床雜志》

作者：

李志輝 , 嚴律南 , 趙永恒 , 劉智敏 , 劉自明 , 李玉

四川大學華西醫院普外科（成都610041）;

關鍵詞：

靶向基因治療腺病毒相關病毒肝癌

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目的構建肝癌特異性HSVTK/GCV重組腺病毒相關病毒質粒并了解其在細胞內的表達。方法以腺病毒相關病毒的質粒WAV2作為載體，將TK基因插在甲胎蛋白（AFP）增強子/白蛋白啟動子(AFP增強子/ALB啟動子)的調控基因的下游，重組成pWAV2/AFPALB/HYTK質粒載體。同時構建質粒載體pEGFP1/AFPALB。然后將上述兩種不同載體分別轉入AFP陽性表達的HepG2細胞株以及陰性表達的7721、SPC 和7901細胞株。結果綠色熒光蛋白只在AFP陽性表達的HepG2細胞株表達，采用PCR技術，以HSVTK的引物擴增所抽提的總DNA中，只有AFP陽性表達的HepG2細胞株擴增出了710 bp的DNA片段。結論pWAV2/AFPALB/HYTK質粒載體在體外實驗中具有很好的靶向性。

引用本文： 李志輝,嚴律南,趙永恒,劉智敏,劉自明,李玉. 人肝癌特異性HSVTK/GCV重組腺病毒相關病毒質粒的構建與研究. 中國普外基礎與臨床雜志, 2004, 11(3): 234-235. doi: 復制

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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. 伍志堅，吳小兵，侯云德. 系列腺病毒伴隨病毒載體的構建及表達β半乳糖苷酶的研究 [Ｊ]. 病毒學報， 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.

《中國普外基礎與臨床雜志》

人肝癌特異性HSVTK/GCV重組腺病毒相關病毒質粒的構建與研究

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《中國普外基礎與臨床雜志》

人肝癌特異性HSVTK/GCV重組腺病毒相關病毒質粒的構建與研究

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

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Content

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