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Repetitive Magnetic Stimulation Modulates the Gene Expression of Neurotrophins and their Receptors in Neuroblastoma
Amanda Stieven1,2, Dirson João Stein1,2, Khetrüin Jordana Fiuza1,2
1Laboratory of Pain Pharmacology and Neuromodulation, Preclinical Investigations, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil.
Background/Aim:
Repetitive magnetic stimulation (rMS) is used to treat neurological conditions. Understanding its modulatory effects requires investigating cellular processes and molecular pathways in vitro. This study aimed to standardize a prototype rMS stimulator-based cell culture method and evaluate tumorigenesis-related cellular and molecular parameters.
Materials And Methods:
Active (75-, 150-, and 300-sec exposure) or sham rMS was administered daily (4 days/300mT-1Hz) to two neuronal [SK-N-BE(2) and SH-SY5Y] and one non-neuronal (HOS) tumor line. Cell viability, cell death, and gene expression of BDNF, NGF, and their receptors, TRKB and TRKA, were evaluated immediately (0 h) and 24 h post-stimulation. Data were analyzed using Kruskal-Wallis/Dunn, one-way ANOVA/Bonferroni, or t-test.
Results:
Both neuroblastoma cell lines, SH-SY5Y (150-sec) and SK-N-BE(2) (75-sec), exhibited increased viability compared to the 300-sec group immediately after treatment; however, none of the stimulated groups was different from sham. rMS increased BDNF, TRKB, NGF, and TRKA gene expression in the 75- and 300-sec groups across both neuroblastoma lines immediately after stimulation. Twenty-four h after rMS, a cell-line and stimulation-time specific reduction in gene expression was observed. Neither cell viability in the three cell lines under investigation, nor cell death in the SK-N-BE(2) were affected by rMS. Twenty-four h after sham or active rMS, all groups demonstrated increased cell viability compared to immediately post-stimulation.
Conclusion:
rMS did not affect cell viability or death in these in vitro models. In the 75- and 300-sec groups, increased neurotrophic gene expression was observed immediately after rMS exposure in both neuroblastoma cell lines, subsequently reducing after 24 h. These data demonstrate molecular effects that are specific to exposure time and cell line, validating this specific in vitro rMS prototype and stimulation protocol.
