• 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(xué)西京醫(yī)院消化病研究所(西安710054);;
  • 3.第四軍醫(yī)大學(xué)西京醫(yī)院肝膽外科(西安710032);

目的　 構(gòu)建受甲胎蛋白（AFP）增強(qiáng)子和白蛋白（Alb）啟動(dòng)子調(diào)控的含有血管抑制素（angiostatin）K5序列的真核表達(dá)載體，通過(guò)基因轉(zhuǎn)染，將angiostatin K5基因?qū)敫伟┘?xì)胞，觀察angiostatin K5的表達(dá)。
方法　 用RTPCR法從正常人真核細(xì)胞擴(kuò)增出angiostatin K5基因，將AFP增強(qiáng)子和Alb啟動(dòng)子調(diào)控序列定向克隆入真核表達(dá)載體pcDNA3.1，并將angiostatin K5 cDNA置于上述調(diào)控序列之下，從而構(gòu)建得到重組真核表達(dá)載體pcDNA3.1AFABangiostatin K5His。利用細(xì)胞培養(yǎng)、脂質(zhì)體介導(dǎo)的細(xì)胞轉(zhuǎn)染，將angiostatin K5基因?qū)敫伟┘?xì)胞。利用SDSPAGE和Western blot法檢測(cè)肝癌細(xì)胞中angiostatin K5的表達(dá)。
結(jié)果 　通過(guò)酶切鑒定及DNA測(cè)序證實(shí)目的真核表達(dá)載體pcDNA3.1AFABangiostatin K5His與預(yù)期的結(jié)果相同。SDSPAGE及Western blot分析證明，angiostatin K5在肝癌細(xì)胞中得以表達(dá)。
結(jié)論 　構(gòu)建的肝癌特異性重組真核表達(dá)載體可增加對(duì)AFP陽(yáng)性肝癌細(xì)胞的治療的特異性，并用于實(shí)現(xiàn)對(duì)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(kāi)宗,竇科峰. 肝癌特異性血管抑制療法的真核表達(dá)載體的構(gòu)建及其表達(dá). 中國(guó)普外基礎(chǔ)與臨床雜志, 2007, 14(1): 15-18. doi: 復(fù)制

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.