• 1.解放軍第451醫(yī)院肝膽普外科(西安710054);;
  • 2.第四軍醫(yī)大學西京醫(yī)院消化病研究所(西安710054);;
  • 3.第四軍醫(yī)大學西京醫(yī)院肝膽外科(西安710032);

目的　 構建受甲胎蛋白（AFP）增強子和白蛋白（Alb）啟動子調控的含有血管抑制素（angiostatin）K5序列的真核表達載體，通過基因轉染，將angiostatin K5基因導入肝癌細胞，觀察angiostatin K5的表達。
方法　 用RTPCR法從正常人真核細胞擴增出angiostatin K5基因，將AFP增強子和Alb啟動子調控序列定向克隆入真核表達載體pcDNA3.1，并將angiostatin K5 cDNA置于上述調控序列之下，從而構建得到重組真核表達載體pcDNA3.1AFABangiostatin K5His。利用細胞培養(yǎng)、脂質體介導的細胞轉染，將angiostatin K5基因導入肝癌細胞。利用SDSPAGE和Western blot法檢測肝癌細胞中angiostatin K5的表達。
結果 　通過酶切鑒定及DNA測序證實目的真核表達載體pcDNA3.1AFABangiostatin K5His與預期的結果相同。SDSPAGE及Western blot分析證明，angiostatin K5在肝癌細胞中得以表達。
結論 　構建的肝癌特異性重組真核表達載體可增加對AFP陽性肝癌細胞的治療的特異性，并用于實現(xiàn)對肝癌的特異性血管抑制作用。

引用本文： 徐宏勇,徐立,高建宏,李開宗,竇科峰. 肝癌特異性血管抑制療法的真核表達載體的構建及其表達. 中國普外基礎與臨床雜志, 2007, 14(1): 15-18. doi: 復制

版權信息： ?四川大學華西醫(yī)院華西期刊社《中國普外基礎與臨床雜志》版權所有，未經授權不得轉載、改編

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.
  1. 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. 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. 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. 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. 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. 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. 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.