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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 investigate the effect of adenovirus-mediated TFPI-2 gene transfection on the invasion of laryngeal squamous cell carcinoma. Methods: Hep-2 cells were transfected with TFPI-2 gene (Ad-TFPI-2) in vitro. The expression of TFPI-2 protein in Hep-2 cells was detected by Western-blot. The expression of Hep-2 Hep-2 cells infected with Ad-TFPI-2 were inoculated subcutaneously in nude mice to observe their tumorigenicity. Results: The correctness of Ad-TFPI-2 was confirmed by enzyme digestion and sequencing. The titer of virus was 2.8 × 1013 PFU / L. Western-blot showed that the expression of TFPI-2 protein was increased in Hep-2 cells Boyden chamber experiments showed that the invasive ability of Hep-2 cells in the experimental group was reduced. The tumorigenicity test in nude mice showed that the ability of Hep-2 cells in the experimental group decreased. Conclusion: TFPI-2 recombinant adenovirus can effectively inhibit the invasion of human laryngeal squamous cell carcinoma Hep-2 cells.