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  

Construction of TFPI-2 recombinant adenovirus and its inhibitive role on invasion of laryngeal squamous carcinoma

作　　者：张慧燕[1] 刘鸣[2] 孙亚男[2] 田霖丽[2] 

机构地区：[1]中国人民解放军第211医院耳鼻咽喉头颈外科,黑龙江哈尔滨150080 [2]哈尔滨医科大学附属第二医院耳鼻咽喉头颈外科,黑龙江哈尔滨150086

出　　处：《中国现代医学杂志》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)

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.

关 键 词：组织途径抑制因子-2基因 腺病毒 喉鳞癌 

分 类 号：R739.63[医药卫生—肿瘤]