Ouyang Jun, Pan Xiaohui, Lin Hui, Hu Zecheng, Xiao Ping, Hu Haobin
Department of Gastrointestinal Surgery, The First Affiliated Hospital of University of South China Hunan, China.
Department of Urology, The First Affiliated Hospital of University of South China Hunan, China.
Am J Transl Res. 2017 Feb 15;9(2):803-811. eCollection 2017.
To investigate the effect of gastric motility protein 2 (GKN2) on the proliferation, apoptosis and invasion of gastric cancer cell and on the JAK/signal transducer and activator of transcription 3 (STAT3) signaling pathway.
Expression of GKN2 was qualified using Western blot analysis in four gastric cancer cell lines and immortalized human gastric mucosal epithelial cell line GES-1. The cells were then transfected with pcDNA3.1-GKN2 and control vector using Lipofectamine2000 and assayed for viability, apoptosis, cell cycle changes, invasion ability as well as expression of cell cycle protein D1 (Cylin D1), Bcl-2, Bax, matrix metalloproteinase 2 (MMP2), MMP9, JAK2 and p-STAT3.
Western blot analyses showed that the expression of GKN2 was significantly lower in 4 gastric cancer cell lines (BGC-823, SGC-7901, AGS and MKN-45) than in GES-1. Of them, SGC-7901 had the lowest expression. The line was chosen for subsequent transfection experiments. Compared with control (transfection with empty vector), pcDNA3.1-GKN2-transfected cells had significantly more GKN2 protein and mRNA, decreased cell viability, increased apoptosis, more cells arrested at G1 phase and reduced invasiveness. Expression analyses showed that expression of Cyclin D1, Bcl-2, MMP2, MMP9, JAK2 and STAT3 was significantly down-regulated, while Bax was significantly up-regulated.
Over-expression of GKN2 can increase apoptosis, reduce proliferation and invasion ability of gastric cancer cells as a result of down-regulated JAK2/STAT3 signaling pathway.
研究胃动蛋白2（GKN2）对胃癌细胞增殖、凋亡、侵袭以及对Janus激酶/信号转导及转录激活因子3（JAK/STAT3）信号通路的影响。
采用蛋白质免疫印迹分析检测4种胃癌细胞系及永生化人胃黏膜上皮细胞系GES-1中GKN2的表达。然后使用Lipofectamine2000将pcDNA3.1-GKN2和对照载体转染至细胞中，并检测细胞活力（细胞活性）、凋亡、细胞周期变化、侵袭能力以及细胞周期蛋白D1（Cyclin D1）、Bcl-2、Bax、基质金属蛋白酶2（MMP2）、MMP9、JAK2和磷酸化STAT3（p-STAT3）的表达。
蛋白质免疫印迹分析显示，4种胃癌细胞系（BGC-823、SGC-7901、AGS和MKN-45）中GKN2的表达明显低于GES-1。其中，SGC-7901的表达最低。选择该细胞系进行后续转染实验。与对照组（转染空载体）相比，转染pcDNA3.1-GKN2的细胞中GKN2蛋白和mRNA明显增多，细胞活力降低，凋亡增加，更多细胞停滞于G1期，侵袭性降低。表达分析显示，Cyclin D1、Bcl-2、MMP2、MMP9、JAK2和STAT3的表达明显下调，而Bax的表达明显上调。
GKN2过表达可通过下调JAK2/STAT3信号通路增加胃癌细胞凋亡，降低其增殖和侵袭能力。
