Glioblastoma multiforme (GBM) presents a significant obstacle in the field of cancer therapy because of its aggressiveness and limited treatment options. Despite advances in surgical resection, radiation therapy, and chemotherapy, the median survival remains dishearteningly brief at 15 months. This review focuses on the mysterious Glioblastoma stem cells (GSCs), particularly those expressing the CD133 marker, acknowledged for their pivotal role in tumor growth and resistance to conventional therapies. The NOTCH signaling pathway emerges as a promising target. This review explores the NOTCH signaling pathway, a crucial regulator of GSCs, and evaluates inhibitors like gamma-secretase inhibitors (GSI), siRNA, and monoclonal antibodies. While recent trials suggest improved outcomes by integrating GSIs with standard treatments, the challenge persists in sparing CD133-positive cells. This review also emphasizes the role of Chk1 and Chk3 inhibitors in the reversal of radioresistance in CD133+ cells. In summary, this review explores the nuances of NOTCH signaling inhibition in GBM treatment, emphasizing precision in targeting the tumor microenvironment and addressing therapeutic resistance associated with CD133-positive cells.