TFPI-2重组腺病毒抑制喉鳞状细胞癌侵袭的实验研究

Inhibiting effects of TFPI-2 recombinant adenoviruson invasi on of laryngeal squamous carcinoma

目的:探讨腺病毒介导的组织因子途径抑制物-2(TFPI-2)基因转染对喉鳞状细胞癌细胞侵袭的影响。方法:将TFPI-2基因(Ad-TFPI-2)体外转染喉鳞状细胞癌Hep-2细胞,运用Western-blot检测Hep-2细胞TFPI-2蛋白表达,运用Boyden小室实验检测Hep-2细胞的侵袭能力的变化,将感染Ad-TFPI-2的Hep-2细胞接种于裸鼠皮下观察其成瘤能力。结果:经酶切、测序等方法的鉴定证实了Ad-TFPI-2的正确性,病毒滴度为2.8×1013PFU/L,Western-blot结果显示TFPI-2蛋白在实验组Hep-2细胞表达增高,Boyden小室实验显示实验组Hep-2细胞的侵袭能力降低,裸鼠成瘤实验显示实验组Hep-2细胞成瘤能力下降。结论:TFPI-2重组腺病毒能够有效地抑制人喉鳞状细胞癌Hep-2细胞的侵袭能力。

Objective：To study the effects of adenovirus-mediated tissue factor pathway inhibitor-2 gene on the invasion of laryngeal squamous cancer. Method： Ad-TFPI-2 was transfected into laryngeal squamous cancer （Hep- 2） cell. Western-blot was used to test the TFPI-2 protein expression and Boyden Chamber experiment was used to examine the invasive ability of Hep-2 cells. Furthermore, the Ad-TFPI-2 infected Hep-2 cells were subcutaneously inoculated in nude mice and the tumor formation capability were observed. Result： Ad-TFPI-2 was identified correctly by endonuclease and sequencing and the virus titer was 2. 8 X 10）a PFU/L. In the Hep-2 cells of treated group, the TFPI-2 protein expression was increased while the invasive capability was descent. The tumor formation capability was also decreased in the treated group nude mouse model. Conelusion：TFPI-2 recombinant adenovirus can effectively inhibit the invasive capability of laryngeal squamous cancer.