Investigating the antiproliferative mechanisms of NaHS, a hydrogen sulfide donor, in the SH-SY5Y cell line

Abstract

Neuroblastoma, a childhood cancer, arises from neural crest cells within the sympathetic nervous system. This study investigated the anticancer effects of the exogenous hydrogen sulfide (H₂S) donor, sodium hydrogen sulfide (NaHS), on neuroblastoma cells, elucidating how it inhibits cell proliferation. The study was conducted using the SH-SY5Y cell line. The cells were exposed to various concentrations of NaHS (32, 16, 8, 4, and 2 mM) for 24 h. Cell viability was assessed using the XTT assay. ELISA was used to measure levels of the growth signal factors HER-2, EGFR, mTOR, ERK-1, and ERK-2, as well as the apoptosis marker caspase-3. In addition, apoptosis was evaluated using the Muse Annexin V & Dead Cell Assay Kit. NaHS significantly reduced cell proliferation, as demonstrated by the XTT assay (p < 0.001). NaHS treatment also decreased levels of HER-2, EGFR, ERK-1, and ERK-2 (p < 0.05 to p < 0.01), while mTOR levels remained unchanged (p > 0.05) and caspase-3 levels increased (p < 0.001). Flow cytometry confirmed that NaHS treatment significantly increased the proportion of apoptotic cells (p < 0.001). The antiproliferative effect of NaHS against neuroblastoma appears to be mediated through the suppression of certain growth signal factors and the induction of apoptosis. These findings suggest that exogenous hydrogen sulfide donors hold promise as a potential therapeutic strategy for neuroblastoma.

Keywords: Apoptosis; EGFR; Hydrogen sulfide; NaHS; SH-SY5Y.

Fig. 1

Effect of NaHS administration on SH-SY5Y cell viability. The data are presented as mean ± SEM. Statistical differences between groups were determined using one-way ANOVA followed by Tukey’s post hoc test. *p < 0.05 and ***p < 0.001 were considered statistically significant compared to the control group

Fig. 2

Effects of NaHS (19.18 mM) administration on cell morphology indicating neuronal damage in SH-SY5Y cells. A Control group (20X), B Control group (40X), C NaHS (19.18 mM) (20X), D NaHS (19.18 mM) (40X) treated group

Fig. 3

The administration of NaHS (19.18 mM) resulted in a reduction of HER-2, EGFR, ERK-1, and ERK-2 levels in SH-SY5Y cells. The quantities of HER-2, EGFR, ERK-1, and ERK-2 were determined using ELISA kits. The data are given as mean ± SEM. Statistical differences between groups were determined using an independent t-test. *p < 0.05, and **p < 0.01 as compared to the control group

Fig. 4

The administration of NaHS (19.18 mM) resulted in an enhancement of Caspase-3 levels in SH-SY5Y cells. The quantity of Caspase-3 was determined using ELISA kits. The data are given as mean ± SEM. Statistical differences between groups were determined using an independent t-test. ***p < 0.001 as compared to the control group

Fig. 5

The administration of NaHS (19.18 mM) affects apoptosis as measured by the Annexin V Binding Assay in SH-SY5Y cells. The data are given as mean ± SEM. Statistical differences between groups were determined using an independent t-test. ***p < 0.001 as compared to the control group