栀子苷增强miR-376b-3p的A-to-Ⅰ编辑抑制糖酵解从而遏制胶质母细胞瘤恶性进展的体内外研究OA

Background and Purpose:Glioblastoma(GBM)is characterized by strong invasiveness,high recurrence rate,and extremely poor prognosis.In-depth exploration of new effective targets and anti-GBM active components holds great significance for improving clinical outcomes.Adenosine-to-inosine(A-to-I)editing is a crucial form of post-transcriptional modification and plays a key role in tumorigenesis and progression.However,its mechanistic role in GBM and its potential as a target for pharmacological intervention remain unclear.Geniposide,the primary active compound extracted from the traditional Chinese medicine Gardenia jasminoides,has shown anti-tumor potential.This study aimed to investigate whether geniposide suppresses GBM progression by regulating A-to-I editing of miR-376b-3p and elucidate the underlying molecular mechanisms.Methods:High-throughput sequencing was used to quantitatively analyze the editing levels of miR-376b-3p in cancer tissues and paired adjacent non-tumor tissues from 32 GBM patients treated at The First Affiliated Hospital of Nanyang Medical College from October 2021 to October 2024.The correlation of these editing levels with the expression of adenosine deaminase acting on RNA type 2(ADAR2)and patient prognosis was further analyzed.In vitro assays evaluated geniposide's effects on GBM cell(U87 and U251)proliferation,migration,invasion and glycolytic metabolism using cell counting kit-8(CCK-8),colony formation assays,EdU cell proliferation assays,Transwell invasion assays,scratch assays,glucose consumption,lactate measurement and cellular energy metabolism analysis.Mechanistic studies involving Western blot,site-directed mutagenesis,and dual-luciferase reporter assays deciphered ADAR2-mediated miRNA editing and target switching.A nude mouse xenograft tumor model was established using BALB/C nude mice(male,4-6 weeks old,n=6 per group,grouped by random number table)to evaluate the in vivo anti-tumor effect of geniposide combined with edited miR-376b-3p agomir.The study was approved by the Ethics Committee of First Affiliated Hospital of Nanyang Medical College(approval number:IRB-Y-L2025139)for clinical samples,and by the Animal Ethics Committee(approval number:IACUC-FJABR-2025022058)for animal experiments.Results:Clinical analysis showed that the A-to-I editing level in GBM was positively correlated with ADAR2 expression(R2=0.466,P<0.001),and patients with high editing had prolonged survival(P=0.028).Geniposide significantly inhibited the proliferation rate of GBM cells,the number of invasive cells,and the healing rate in scratch assays by reprogramming glycolytic metabolism GBM cells(P<0.001).Its mechanism is to specifically upregulate ADAR2 expression,thereby promoting A-to-I editing of miR-376b-3p.Mechanistically,A-to-I editing switched miR-376b-3p targeting from the tumor suppressor CBX7 to the oncogene TMX2(dual luciferase assay confirmed altered binding,P<0.001),leading to inhibition of the key glycolysis pathway.TMX2 overexpression reversed the anti-tumor effects of geniposide combined with edited miR-376b-3p.In vivo experiments further confirmed that the combination therapy can most effectively inhibit tumor growth(P<0.001)and downregulate the expression of hexokinase 2(HK2),pyruvate kinase M2(PKM2),and glucose transporter 1(GLUT1).Conclusion:Geniposide activates ADAR2-mediated A-to-I editing of miR-376b-3p,redirecting its targeting from CBX7 to TMX2 to inhibit glycolytic reprogramming and GBM progression.These findings may provide a novel clue for developing RNA editing-based therapies for GBM.