Differential Expression of Tyrosine Hydroxylase Protein and Apoptosis-Related Genes in Differentiated and Undifferentiated SH-SY5Y Neuroblastoma Cells Treated with MPP⁺

Abstract

The human neuroblastoma SH-SY5Y cell line has been used as a dopaminergic cell model for Parkinson's disease research. Whether undifferentiated or differentiated SH-SY5Y cells are more suitable remains controversial. This study aims to evaluate the expression of apoptosis-related mRNAs activated by MPP+ and evaluate the differential expression of tyrosine hydroxylase (TH) in undifferentiated and retinoic acid- (RA-) induced differentiated cells. The western blot results showed a gradual decrease in TH in undifferentiated cells and a gradual increase in TH in differentiated cells from days 4 to 10 after cell plating. Immunostaining revealed a gradual increase in TH along with neuritic outgrowth in differentiated cells on days 4 and 7 of RA treatment. For the study on cell susceptibility to MPP+ and the expression of apoptosis-related genes, MTT assay showed a decrease in cell viability to approximately 50% requiring 500 and 1000 μM of MPP+ for undifferentiated and RA-differentiated cells, respectively. Using real-time RT-PCR, treatment with 500 μM MPP+ led to significant increases in the Bax/Bcl-2 ratio, p53, and caspase-3 in undifferentiated cells but was without significance in differentiated cells. In conclusion, differentiated cells may be more suitable, and the shorter duration of RA differentiation may make the SH-SY5Y cell model more accessible.

Differentiation enhances Zika virus infection of neuronal brain cells

“…Following treatment with RA, there was an increase in the expression of multiple established markers of neuronal differentiation (Table 1 and Supplementary Fig. 3B ), including MAP2 (microtubule-associated protein-2), synaptophysin (present at functional synapses), SatB (a transcriptional factor expressed in cortical neurons), GFAP (a marker of astrocytes), and TH (tyrosine hydroxyolase, a dopaminergic marker) 34 , 35 , as well as corresponding expected decreases in the abundance of vimentin and ID2 genes (Table 1 ) 36 .…”

Tectorigenin attenuates the MPP+‑induced SH‑SY5Y cell damage, indicating a potential beneficial role in Parkinson's disease by oxidative stress inhibition

“…In the 1980s, MPTP was reported to cause Parkinsonian symptoms (21), and it was later observed that it is biotransformed into toxic MPP + by glial cells, which can cross the blood-brain barrier and enter dopaminergic neurons, resulting in mitochondrial damage, the generation of oxidative stress and the activation of pro-apoptotic pathways (22). In agreement with these previous studies, the present study observed that treatment of SH-SY5Y cells with MPP + induced loss of cell viability, occurrence of apoptotic features, enhancement of caspase-3 activity and cytochrome c, which was associated with the increases in the ratio of Bax/Bcl-2 and the elevation of ROS levels in SH-SY5Y cells.…”

Echinacoside Protects Against MPP+-Induced Neuronal Apoptosis via ROS/ATF3/CHOP Pathway Regulation

“…-induced ATF3 and CHOP expression in neuroblastoma cells [9,23]. ATF3 upregulates p53 expression and induces apoptosis in SH-SY5Y cells [23] and other cells [17,36]. Interestingly, a recent study showed that ATF3 upregulation of p53 plays an important role in apoptosis induced by MPP ?…”

“…Interestingly, a recent study showed that ATF3 upregulation of p53 plays an important role in apoptosis induced by MPP ? [36]. CHOP was first identified as a transcription factor that is induced in response to growth arrest and DNA damage.…”