Construction and Expression of Hepatocellular CarcinomaSpecific Expressing Eukaryotic Vector for AntiAngiogenesis Therapy_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

XU Hongyong , XU Li , GAO Jianhong , LI Kaizong , DOU Kefeng

Hepatobiliary General Surgery， The 451th Hospital of PLA， Xi’an 710054, China;

Corresponding author：

Keywords：

Hepatocellular carcinoma; Angiostatin; αFetoprotein enhancer

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Objective　 To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of αfetoprotein （AFP） enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection.
Methods 　Angiostatin cDNA was amplified from normal human eukaryotic cells by using RTPCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1AFABangiostatin K5His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with antiHis antibody.
Results　 The 500base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1AFABangiostatin K5His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDSPAGE and Western blot.
Conclusion　 Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.

Citation： XU Hongyong,XU Li,GAO Jianhong,LI Kaizong,DOU Kefeng. Construction and Expression of Hepatocellular CarcinomaSpecific Expressing Eukaryotic Vector for AntiAngiogenesis Therapy. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2007, 14(1): 15-18. doi: Copy

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1. 　Farinati F, Marino D, De Giorgio M, et al. Diagnostic and prognostic role of alphafetoprotein in hepatocellular carcinoma: both or neither? [J]. Am J Gastroenterol, 2006; 101(3)∶524.
2. 　Perri SR, Nalbantoglu J, Annabi B, et al. Plasminogen kringle 5engineered glioma cells block migration of tumorassociated macrophages and suppress tumor vascularization and progression [J]. Cancer Res, 2005; 65(18)∶8359.
3. 　Kong N, Lim D, Lee K. Interacting partners for kringle domains of plasminogen: common binding with K1 and K5 domains [J]. Protein Pept Lett, 2004; 11(6)∶521.
4. 　Su H, Lu R, Chang JC, et al. Tissuespecific expression of herpes simplex virus thymidine kinase gene delivered by adenoassociated virus inhibits the growth of human hepatocellular carcinoma in athymic mice [J]. Proc Natl Acad Sci USA, 1997; 94(25)∶13891.
5. 　Cao YH, Andrew Chen, Seong SA, et al. Kringle5 of plasminogen is a novel inhibitor of endothelial cell growth [J]. J Biol Chem, 1997; 272(36)∶22924.
6. 　Su H, Chang JC, Xu SM, et al. Selective killing of AFPpositive hepatocellular carcinoma cells by adenoassociated virus transfer of the herpes simplex virus thymidine kinase gene [J]. Hum Gene Ther, 1996; 7(4)∶463.
7. 　Berent R, Auer J, Porodko M, et al. Influence of cardiopulmonary resuscitation on levels of tumour markers [J]. Eur J Cancer Care (Engl), 2006; 15(3)∶252.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction and Expression of Hepatocellular CarcinomaSpecific Expressing Eukaryotic Vector for AntiAngiogenesis Therapy

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