siRNA targeting of Trop2 suppresses the proliferation and invasion of lung adenocarcinoma H460 cells
siRNA targeting of Trop2 suppresses the proliferation and invasion of lung adenocarcinoma H460 cells
复制标题
DOI：
10.3892/etm.2015.2530
复制
发表时间：
2015-08-01
影响因子：
2.7
通讯作者：
Jiang, Ai-Gui
中科院分区：
文献类型：
--
作者：
Gao, Xiao-Yan;Zhu, Ye-Han;Jiang, Ai-Gui
本研究旨在探讨小干扰RNA（siRNA）抑制Trop 2基因对肺腺癌H460细胞增殖和侵袭能力的影响。将靶向Trop 2开放阅读框的siRNA重组腺病毒表达载体（rAd 5-siTrop 2）转染肺腺癌H460细胞。研究中包括三组，即对照组（未转染对照）、rAd 5-siCtrl组（天然对照）和rAd 5-siTrop 2组（敲低Trop 2基因）。采用定量聚合酶链反应（PCR）和蛋白质印迹法（Western blot）分别检测Trop 2 mRNA和蛋白表达水平。Western blot检测细胞周期蛋白D1（cyclin D1）和磷酸化细胞外信号调节激酶（p-ERK）-1（p-ERK-1）的表达。采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化物和Transwell法研究Trop 2抑制剂对肺腺癌H460细胞增殖和侵袭的影响。成功构建了靶向Trop 2的siRNA重组质粒。重组腺病毒载体rAd 5-siTrop 2可显著下调肺腺癌H460细胞中Trop 2 mRNA和蛋白的表达，同时下调cyclin D1和p-ERK-1的表达。此外，Trop 2基因沉默后，肺腺癌H460细胞的增殖和侵袭率降低。因此，Trop 2在肺腺癌H460细胞的体外增殖和侵袭中起关键作用。
The aim of the present study was to investigate the effect of the small interfering RNA (siRNA)-induced inhibition of the Trop2 gene on the proliferation and invasion of lung adenocarcinoma H460 cells. A recombinant adenovirus expression vector, which contained siRNA targeting open reading frames for Trop2 (rAd5-siTrop2), was transfected into lung adenocarcinoma H460 cells. Three groups were included in the study, namely the Ctrl (non-transfected control), rAd5-siCtrl (native control) and rAd5-siTrop2 (knockdown Trop2 gene) groups. The mRNA and protein expression levels of Trop2 were detected using quantitative polymerase chain reaction and western blot analysis, respectively. In addition, the expression levels of cyclin Dl and phospho-extracellular signal regulated kinase (p-ERK)-1 were detected using western blot analysis. The effects of Trop2 inhibition on the proliferation and invasion of lung adenocarcinoma H460 cells were investigated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assay. Trop2-targeted siRNA recombinant plasmids were successfully constructed. The recombinant adenovirus vector, rAd5-siTrop2, significantly downregulated the mRNA and protein expression levels of Trop2 in the lung adenocarcinoma H460 cells, with cyclin D1 and p-ERK-1 expression downregulated simultaneously. In addition, following the silencing of Trop2, the proliferation and invasion rates of the lung adenocarcinoma H460 cells were reduced. Therefore, the results indicated that Trop2 serves a key function in the proliferation and invasion of lung adenocarcinoma H460 cells in vitro.