Zhou Lingshan, Yang Yuan, Qiao Qian, Mi Yingying, Gan Yuling, Zheng Ya, Wang Yuping, Liu Min, Zhou Yongning
The First Clinical Medical College, Lanzhou University, Lanzhou, 730000, China.
Department of Gastroenterology, The First Hospital of Lanzhou University, Lanzhou, 730000, China.
J Cancer Res Clin Oncol. 2024 Dec 26;151(1):15. doi: 10.1007/s00432-024-06039-z.
This study aimed to investigate that AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer (GC) progression via targeted regulation of IRS-1 transcription.
The study utilized databases such as PhosphositePlus, TRANSFAC, CHEA, GPS 5.0, and TCGA, along with experimental techniques including Western Blot, co-IP, in vitro kinase assay, construction of lentiviral overexpression and silencing vectors, immunoprecipitation, modified proteomics, immunofluorescence, ChIP-PCR, EdU assay, Transwell assay, and scratch assay to investigate the effects of AKT1-induced Notch1 phosphorylation on cell proliferation, invasion and migration in vitro, as well as growth and epithelial-mesenchymal transition (EMT) in vivo.
AKT1 was found to induce phosphorylation of Notch1 at the S2183 site in GC, subsequently altering the subcellular localization of Notch1-IC and promoting its nuclear translocation. The transcription factor RBPJ that binds to Notch1 transcriptionally regulated IRS-1, CDH5, TNL1, ASCL2, and LRP6. Experimental validation revealed that Notch1-IC can regulate the expression of IRS-1. Overexpression of Notch1-IC was shown to promote the proliferation, invasion, and metastasis of GC cells, while knockdown of IRS-1 partially inhibited the aforementioned effects induced by Notch1-IC overexpression. Further experiments in vitro and vivo confirmed that AKT1-induced Notch1 phosphorylation can regulate the expression of IRS-1 and promote the malignant behavior of GC, including proliferation, invasion, metastasis, and EMT, with knockdown of IRS-1 partially reversing these effects.
AKT1 induces the Notch1 phosphorylation and promotes the activation and nuclear translocation of Notch1-IC by targeting the regulation of IRS-1, thereby advancing the progression of GC.
本研究旨在探讨AKT1介导的NOTCH1磷酸化通过靶向调控IRS-1转录促进胃癌（GC）进展。
本研究利用了PhosphositePlus、TRANSFAC、CHEA、GPS 5.0和TCGA等数据库，以及蛋白质免疫印迹、免疫共沉淀、体外激酶分析、慢病毒过表达和沉默载体构建、免疫沉淀、修饰蛋白质组学、免疫荧光、染色质免疫沉淀PCR、EdU分析、Transwell分析和划痕分析等实验技术，以研究AKT1诱导的Notch1磷酸化对体外细胞增殖、侵袭和迁移以及体内生长和上皮-间质转化（EMT）的影响。
发现AKT1在GC中诱导Notch1在S2183位点磷酸化，随后改变Notch1-IC的亚细胞定位并促进其核转位。与Notch1结合的转录因子RBPJ转录调控IRS-1、CDH5、TNL1、ASCL2和LRP6。实验验证表明Notch1-IC可调控IRS-1的表达。Notch1-IC的过表达促进GC细胞的增殖、侵袭和转移，而IRS-1的敲低部分抑制了Notch1-IC过表达诱导的上述效应。进一步的体外和体内实验证实，AKT1诱导的Notch1磷酸化可调控IRS-1的表达并促进GC的恶性行为，包括增殖、侵袭、转移和EMT，IRS-1的敲低可部分逆转这些效应。
AKT1通过靶向调控IRS-1诱导Notch1磷酸化并促进Notch1-IC的激活和核转位，从而推动GC进展。
