CHRM2 drives neuroblastoma progression via the PI3K/AKT pathway: mechanistic insights and Genistein-based therapeutic intervention

Abstract

Neuroblastoma (NB) remains a challenging pediatric malignancy with limited therapeutic targets. While PI3K/AKT signaling is implicated in NB progression, novel regulators within this pathway require exploration. This study identifies CHRM2 as a critical mediator and investigates the therapeutic potential of Genistein (GST) in modulating CHRM2-driven NB pathogenesis. Integrated bioinformatics analysis identified key genes linked to PI3K/AKT signaling. CHRM2 was prioritized for functional validation using PCR, western blot, CCK-8, TUNEL, wound healing, transwell, and ELISA in SH-SY5Y cells. Gain- and loss-of-function experiments (CHRM2, si-CHRM2) and PI3K/AKT modulators (LY294002, 740 Y-P) were employed to dissect mechanistic interactions. Bioinformatics analysis revealed CHRM2 as a consistently upregulated gene in advanced-stage NB patients and a PI3K/AKT-associated hub gene. GST treatment suppressed SH-SY5Y cell viability and reduced CHRM2 expression, correlating with diminished LDH/NSE release. CHRM2 knockdown inhibited proliferation, migration, and survival while promoting apoptosis, whereas its overexpression exerted opposing effects. GST synergized with CHRM2 knockdown to amplify anti-tumor outcomes. Mechanistically, CHRM2 ablation enhanced GST-induced apoptosis via caspase 3/7 and cytochrome C activation, while PI3K/AKT pathway activation (740 Y-P) reversed GST effects. Conversely, PI3K/AKT inhibition (LY294002) exacerbated GST-mediated apoptosis and functional suppression, which was attenuated by CHRM2 overexpression. CHRM2 drives NB progression by sustaining PI3K/AKT signaling, and GST exerts anti-tumor effects via CHRM2 suppression. These findings nominate CHRM2 as a novel therapeutic target and highlight GST as a promising agent for NB treatment through PI3K/AKT pathway modulation.