Related Experiment Video

Updated: May 28, 2026

A Rapid Screening Workflow to Identify Potential Combination Therapy for GBM using Patient-Derived Glioma Stem Cells

A Rapid Screening Workflow to Identify Potential Combination Therapy for GBM using Patient-Derived Glioma Stem Cells

Published on: March 28, 2021

The Flavonoid Rutin Enhances Temozolomide Sensitivity in Glioblastoma Spheroids by Modulating Chemoresistance via

Irlã Santos Lima¹, Fernanda Vidal Carvalho¹, Érica Novaes Soares¹

¹Laboratory of Neurochemistry and Cellular Biology (LabNq), Institute of Health Sciences, Federal University of Bahia, Salvador 40231-300, BA, Brazil.

Antioxidants (Basel, Switzerland)

|May 27, 2026

Abstract

Introduction:

Glioblastoma (GBM) is the most aggressive primary tumor of the central nervous system and is highly resistant to temozolomide (TMZ). Rutin is a potent antioxidant with immunomodulatory and anti-glioma effects in vitro, although its mechanisms of action remain incompletely understood. This study investigated the effects of rutin on morphology, viability, redox balance, and pro-tumoral signaling in GBM 2D cultures and 3D spheroids, as well as its association with TMZ sensitivity.

Methods:

GL15 and U343 human GBM cell lines and primary astrocytes were treated with rutin (5-30 μM) and/or TMZ (125-4000 μM). Cell metabolic activity and viability were assessed by MTT, PI/DiOC18(3) or PI/Hoechst. Cell migration was assessed from spheroid-derived cells, and extracellular matrix (ECM) components (fibronectin and laminin) were evaluated by immunofluorescence. Intracellular reactive oxygen species (ROS) were measured by DCFH-DA fluorescence. IL-6, STAT3, NOS2, and IDO1 gene expression were determined by RT-qPCR, and protein expression of MMP2, fibronectin, STAT3, PI3K, and AKT by Western blotting. Nitric oxide (NO) and L-kynurenine levels were quantified in the supernatant by colorimetric assays.

Results:

Rutin reduced cell viability and enhanced TMZ cytotoxicity in both 2D and 3D cultures, while exerting selective effects by increasing metabolic activity and attenuating TMZ-induced effects in non-tumoral primary astrocytes. In 3D spheroids, rutin affected structural organization and reduced spheroid-derived cell migration, accompanied by changes in ECM components, including MMP2, fibronectin, and laminin. Rutin decreased intracellular ROS levels and suppressed the TMZ-induced increase in ROS and NOS signaling. These effects were accompanied by modulation of IL-6/STAT3 signaling, along with reduced STAT3, PI3K, and AKT protein levels. Rutin also modulated immunometabolic parameters, including extracellular L-kynurenine and nitric oxide levels, and enhanced TMZ responsiveness following pre-sensitization.

Conclusions:

Rutin enhances TMZ responsiveness by modulating interconnected pro-tumoral mechanisms, including redox balance, pro-survival signaling, ECM remodeling and migratory behavior, and immunometabolic pathways linked to chemoresistance, supporting its potential as an adjuvant therapeutic strategy.

Insights

Rutin enhances glioblastoma (GBM) treatment by improving temozolomide (TMZ) effectiveness. This natural compound modulates redox balance, signaling pathways, and cell migration, supporting its use as an adjuvant therapy.

Area of Science:

Oncology
Pharmacology
Biochemistry

Background:

Glioblastoma (GBM) is a highly aggressive brain tumor resistant to temozolomide (TMZ).
Rutin, a natural antioxidant, exhibits anti-glioma properties, but its mechanisms are not fully understood.
This study explores rutin's effects on GBM and its synergy with TMZ.

Purpose of the Study:

Investigate rutin's impact on GBM cell morphology, viability, and redox balance.
Determine rutin's influence on pro-tumoral signaling pathways in GBM.
Assess rutin's potential to enhance temozolomide (TMZ) sensitivity in glioblastoma.

Main Methods:

Utilized human GBM cell lines (GL15, U343) and primary astrocytes.
Assessed cell viability, metabolic activity, and migration.
Measured reactive oxygen species (ROS), nitric oxide (NO), and L-kynurenine.
Analyzed gene and protein expression of key signaling molecules (e.g., STAT3, PI3K, AKT, MMP2).

Main Results:

Rutin reduced GBM cell viability and increased TMZ cytotoxicity in 2D and 3D cultures.
Rutin modulated extracellular matrix components, decreased ROS, and suppressed NOS signaling.
Rutin inhibited IL-6/STAT3 and PI3K/AKT pathways and altered immunometabolic parameters.
Rutin demonstrated selective effects, enhancing astrocyte viability while impacting GBM cells.

Conclusions:

Rutin enhances TMZ responsiveness by targeting multiple pro-tumoral mechanisms.
Rutin modulates redox balance, signaling, migration, and immunometabolism in GBM.
Rutin shows potential as an adjuvant therapy to improve glioblastoma treatment outcomes.

Read full analysis

Keywords:

PI3K/AKT STAT3 chemoresistance extracellular matrix remodeling glioblastoma kynurenine metabolism microtumors oxidative stress rutin temozolomide

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