Chen Jia, Zhang Shoude, Lin Yi, Yang Beibei, Cao Jiang
Department of Otorhinolaryngology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, 88 Jiefang Road, Hangzhou, Zhejiang Province, 310009, People's Republic of China.
Department of Otorhinolaryngology,Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, 310016, People's Republic of China.
Pathol Oncol Res. 2017 Oct;23(4):829-835. doi: 10.1007/s12253-016-0160-8. Epub 2017 Jan 18.
The purpose of this study was to develop a recombinant adenovirus with secretory leukoprotease inhibitor (SLPI) promoter-controlled expression for gene therapy of squamous cell carcinoma (SCC). An artificial microRNA targeting epidermal growth factor receptor (EGFR) was designed, and used to construct a replication-defective recombinant adenovirus with SLPI promoter-controlled expression. The silencing efficiency of this vector (Ad-SLPI-EGFRamiR) was detected in Hep-2 cells. Western blotting showed that the expression of 170 kD EGFR was significantly reduced in Hep-2 cells 72 h after infection with Ad-SLPI-EGFRamiR. At a multiplicity of infection (MOI) of 200 pfu/cell, proliferation of Hep-2 cells was highly inhibited by Ad-SLPI-EGFRamiR (inhibition rate: ~70%). The apoptosis rate of Hep-2 cells at 72 h after infection with Ad-SLPI-EGFRamiR at a MOI 35 pfu/cell was 32.8%. The adenovirus constructed was able to specifically inhibit the growth of SCC cells in vitro.
本研究的目的是构建一种由分泌型白细胞蛋白酶抑制剂（SLPI）启动子控制表达的重组腺病毒，用于鳞状细胞癌（SCC）的基因治疗。设计了一种靶向表皮生长因子受体（EGFR）的人工微小RNA，并用于构建具有SLPI启动子控制表达的复制缺陷型重组腺病毒。在Hep-2细胞中检测了该载体（Ad-SLPI-EGFRamiR）的沉默效率。蛋白质印迹法显示，用Ad-SLPI-EGFRamiR感染Hep-2细胞72小时后，170 kD EGFR的表达显著降低。在感染复数（MOI）为200 pfu/细胞时，Ad-SLPI-EGFRamiR对Hep-2细胞的增殖有高度抑制作用（抑制率：约70%）。在MOI为35 pfu/细胞时，用Ad-SLPI-EGFRamiR感染Hep-2细胞72小时后的凋亡率为32.8%。构建的腺病毒能够在体外特异性抑制SCC细胞的生长。
