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组织途径抑制因子-2基因重组腺病毒载体的构建及其抑制喉鳞癌侵袭的研究 被引量：2

Construction of TFPI-2 recombinant adenovirus and its inhibitive role on invasion of laryngeal squamous carcinoma
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摘要 目的构建载有TFPI-2基因的重组腺病毒载体,探讨其对喉癌侵袭能力的抑制作用。方法将TFPI-2cDNA克隆于腺病毒载体PSG-CMV,得到重组质粒PSG-TFPI-2。将质粒PSG-TFPI-2与5型腺病毒共转染至293细胞,生成带有TFPI-2基因的重组腺病毒载体。重组腺病毒质粒经过293细胞的扩增和CsCl的纯化,感染Hep-2细胞。运用侵袭实验观察感染后的喉癌细胞侵袭能力的变化。结果经酶切和测序等方法的鉴定,证实了载体构建的正确性,病毒滴度为2.8×1010PFU/mL。侵袭实验显示转染Ad-TFPI-2的喉癌细胞侵袭能力下降。结论成功构建带有TFPI-2的重组腺病毒载体,运用Ad-TFPI-2重组腺病毒能够有效地抑制喉癌细胞的侵袭能力。 [ Objectives ] To construct tissue factor pathway inhibitor-2 recombinant adenovirus （Ad-TFPI-2） and study its effect on the invasion of laryngeal squamous cancer. [Methods] TFPI-2 eDNA was cloned into PSG-CMV vector to gain PSG-TFPI-2. PSG-TFPI-2 and 5-type adenovirus were transfeeted into 293 cells and the recombinant adenovirus carrying TFPI-2 gene was created. The recombinant adenovirus was transfected into laryngeal squamous cancer （Hep-2） cell after amplificated in 293 cells and purified in CsCI. Boyden assay was used to observe the invasive capility of Hep-2 cells. [Results] Ad-TFPI-2 was identified correctly by endonuclease and sequencing and the virus titer was 2.8×10^10 PFU/mL. Boyden assay showed the invasive capability in the Hep-2 cells of treated group was descent. [ Conclusion ] TFPI-2 recombinant adenovirus was constructed successfully and it can effectively inhibit the invasive capability of laryngeal squamous cancer.
作者 张慧燕 刘鸣 孙亚男 田霖丽
机构地区 中国人民解放军第 哈尔滨医科大学附属第二医院耳鼻咽喉头颈外科
出处 《中国现代医学杂志》 CAS CSCD 北大核心 2009年第18期2741-2743,共3页 China Journal of Modern Medicine
基金 黑龙江省攻关课题(No:GC07C35106)
关键词 组织途径抑制因子-2基因 腺病毒 喉鳞癌 adenovirus tissue factor pathway inhibitor-2 gene therapy laryngeal squamous carcinoma
分类号 R739.63 [医药卫生—肿瘤]
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参考文献9

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二级参考文献4

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共引文献6

同被引文献36

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中国现代医学杂志
2009年 第18期

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