title: 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.
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Kawinthra
authors.initials: K
authors.lastname: Khwanraj
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand ; Department of Pharmacology, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Chareerut
authors.initials: C
authors.lastname: Phruksaniyom
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Suriyat
authors.initials: S
authors.lastname: Madlah
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Permphan
authors.initials: P
authors.lastname: Dharmasaroja