• 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）增強子和白蛋白（Alb）啟動子調(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)胞擴增出angiostatin K5基因，將AFP增強子和Alb啟動子調(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法檢測肝癌細(xì)胞中angiostatin K5的表達(dá)。
結(jié)果 　通過酶切鑒定及DNA測序證實目的真核表達(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AFP陽性肝癌細(xì)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(dá)載體的構(gòu)建及其表達(dá). 中國普外基礎(chǔ)與臨床雜志, 2007, 14(1): 15-18. doi: 復(fù)制

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