Department of Emergency Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People's Republic of China.
Department of General Surgery, First Affiliated Hospital of Anhui Medical University, 218 JiXi Avenue, Hefei, 230022, People's Republic of China.
Mol Cell Biochem. 2018 Jun;443(1-2):93-100. doi: 10.1007/s11010-017-3213-8. Epub 2017 Oct 25.
The purpose of the article is to investigate the role of IARS2 in proliferation, apoptosis, and cell cycle of gastric cancer (GC) cells in vitro. The IARS2-shRNA lentiviral vector was established and used to infect the GC cell line AGS. qRT-PCR and Western blot were employed to determine the efficiency of IARS2 knockdown. The effects of IARS2 knockdown on cell proliferation, cell clone formation, and cell cycle were assessed by MTT assay, colony formation assay, and flow cytometer analysis, respectively. Finally, a PathScan Antibody Array Kit was used to detect the expression levels of cell cycle-related proteins after IARS2 knockdown in AGS cells to elucidate the underlying mechanisms. Compared with negative control group, IARS2 was significantly knocked down by transfection with lentivirus encoding shRNA of IARS2 in AGS cells. IARS2 knockdown significantly inhibited the proliferation and colony formation ability and induced cycle arrest at G2/M phase of AGS cells. IARS2 knockdown significantly decreased the expression levels of phosphorylation of (p-Smad2), p-SAPK/JUK, cleavage-Caspase-7, and p-TAK1, but increased the expression levels of p-53 and cleavage-PARP in AGS cells compared to shCtrl group. We demonstrated that IARS2 knockdown inhibits proliferation, suppresses colony formation, and causes cell cycle arrest in AGS cells. We also found that IARS2 regulates key molecules of cell apoptosis-related signaling pathway.
本文旨在研究 IARS2 在体外对胃癌（GC）细胞增殖、凋亡和细胞周期的作用。构建了 IARS2-shRNA 慢病毒载体，并用于感染 GC 细胞系 AGS。采用 qRT-PCR 和 Western blot 检测 IARS2 敲低效率。通过 MTT 检测、集落形成实验和流式细胞仪分析分别评估 IARS2 敲低对细胞增殖、细胞克隆形成和细胞周期的影响。最后，使用 PathScan 抗体阵列试剂盒检测 IARS2 敲低后 AGS 细胞中与细胞周期相关蛋白的表达水平，以阐明其潜在机制。与阴性对照组相比，AGS 细胞中 IARS2 经 shRNA 慢病毒转染后表达明显下调。IARS2 敲低显著抑制 AGS 细胞的增殖和集落形成能力，并诱导细胞周期停滞在 G2/M 期。与 shCtrl 组相比，IARS2 敲低后 AGS 细胞中磷酸化 Smad2（p-Smad2）、SAPK/JUK 磷酸化（p-SAPK/JUK）、Caspase-7 裂解、TAK1 磷酸化（p-TAK1）的表达水平降低，p53 和 PARP 裂解（cleavage-PARP）的表达水平升高。我们证明了 IARS2 敲低抑制 AGS 细胞增殖、抑制集落形成并导致细胞周期停滞。我们还发现 IARS2 调节细胞凋亡相关信号通路的关键分子。
