Haloxylon scoparium pomel fractions suppress neurosphere formation and induce apoptosis in U87 glioblastoma

Haloxylon scoparium Pomel (HSP) is a medicinal plant, with its therapeutic applications widely recognized across North African populations. Although HSP is acknowledged for its curative properties, its antitumor potential remains largely unexplored. This study addresses this gap by evaluating the anticancer activity of HSP fractions on human U87 glioblastoma cells cultured as two-dimensional (2D) monolayers and three-dimensional (3D) neurospheres. Using MTT assays, we determined the IC₅₀ values for ethyl acetate and aqueous fractions, revealing the unexpected finding that the butanolic fraction exhibited no effect on cell viability. In vitro wound-healing assays confirmed a concentration-dependent antiproliferative effect of the ethyl acetate fraction from both ethanolic (EAFEE) and methanolic (EAFME) extracts, implicating modulation of key migration biomarkers such as Fibronectin (FN) and MT1-MMP. Notably, these fractions also significantly inhibited 3D neurosphere formation. Flow cytometry analysis of cell cycle distribution supported the antiproliferative activity, accompanied by reduced expression of cell cycle regulators. Furthermore, Annexin V assays demonstrated a marked increase in apoptosis following treatment with EAF fractions. Gene expression profiling of treated neurospheres and monolayers revealed differential regulation of pro-apoptotic and pro-inflammatory signatures. Collectively, these findings provide evidence that HSP EAFEE can disrupt invasive, apoptotic, and inflammatory pathways implicated in glioblastoma progression, underscoring its potential as a novel candidate for anticancer therapy.