Notch intracellular domain regulates glioblastoma proliferation through the Notch1 signaling pathway
Yixuan Wang, Qian Sun, Rongxin Geng, Hao Liu, Fanen Yuan, Yang Xu, Yangzhi Qi, Hongxiang Jiang, Qianxue Chen, Baohui Liu
Abstract
Notch intracellular domain (NICD), also known as the activated form of Notch1 is closely associated with cell differentiation and tumor invasion. However, the role of NICD in glioblastoma (GBM) proliferation and the underlying regulatory mechanism remains unclear. The present study aimed to investigate the expression of NICD and Notch1 downstream gene HES5 in human GBM and normal brain samples and to further detect the effect of NICD on human GBM cell proliferation. For this purpose, western blotting and immunohistochemical staining were performed to analyze the expression of NICD in human GBM tissues, while western blotting and reverse‑transcription quantitative PCR experiments were used to analyze the expression of Hes5 in human GBM tissues. A Flag‑NICD vector was used to overexpress NICD in U87 cells and compound E and small interfering (si) Notch1 were used to downregulate NICD. Cellular proliferation curves were generated and BrdU assays performed to evaluate the proliferation of U87 cells. The results demonstrated that compared with normal brain tissues, the level of NICD protein in human GBM tissues was upregulated and the protein and mRNA levels of Hes5 were also upregulated in GBM tissues indicating that the Notch1 signaling pathway is activated in GBM. Overexpression of NICD promoted the proliferation of U87 cells <em>in vitro</em> while downregulation of NICD by treatment with compound E or siNotch1 suppressed the proliferation of U87 cells <em>in vitro</em>. In conclusion, NICD was upregulated in human GBM and NICD promoted GBM proliferation via the Notch1 signaling pathway. NICD may be a potential diagnostic marker and therapeutic target for GBM treatment.