Investigation of the effects of a sulfite molecule on human neuroblastoma cells via a novel oncogene URG4/URGCP.
OBJECTIVE
The aim of this study is to determine the anticancer effect of sulfite on SH-SY5Y neuroblastoma cells in vitro conditions and elucidate underlying molecular mechanism of sulfite and explore its therapeutic activity.


METHODS
In this study, cytotoxic effects of sulfite in SH-SY5Y cels were detected over time in a dose dependent manner with the IC50 doses ranging from 0.5 to 10 mM. Genotoxic effect of sulfite was shown by comet assay. IC50 doses in the SH-SY5Y cells were detected as 5 mM. Expression profiles of the target genes related to apoptosis and cell cycle control were determined by quantitative RT-PCR. Protein changes were determined by western blot analysis.


RESULTS
URG4/URGCP, CCND1, CCND2, CDK4, CDK6, E2F4 and BCL-2 gene expression levels were significantly reduced and RB1, TP53, BAX, BID, CASP2, CASP3, CASP9 and DIABLO gene expressions were significantly increased in dose group cells. The mechanism of this result may be related to sulfite dependent inhibition of cell cycle at the G1 phase by down-regulating URG4/URGCP or CCND1, CDK4, CDK6 gene expression and stimulating apoptosis via the intrinsic pathway. Sulfite suppressed invasion and colony formation in SH-SY5Y cell line using matrigel invasion chamber and colony formation assay, respectively.


CONCLUSIONS
It is thought that sulfite demonstrates anticarcinogenesis activity by affecting cell cycle arrest, apoptosis s, invasion, and colony formation on SH-SY5Y cells. Sulfite may be an effective agent for treatment of neuroblastoma as a single agent or in combination with other agents.
Pamukkale University School of Medicine, Department of Medical Biology, Denizli, Turkey. Electronic address: yavuzdodurga@gmail.com.
Yavuz
Y
Dodurga
Pamukkale University School of Medicine, Department of Medical Biology, Denizli, Turkey.
Mücahit
M
Seçme
Pamukkale University School of Medicine, Department of Medical Biology, Denizli, Turkey.
Canan
C
Eroğlu
Pamukkale University School of Medicine, Department of Physiology, Denizli, Turkey.
Gülşah
G
Gündoğdu
Ege University School of Medicine, Department of Medical Biology, İzmir, Turkey.
Çığır Biray
ÇB
Avcı
Pamukkale University School of Medicine, Department of Medical Biology, Denizli, Turkey.
Gülseren
G
Bağcı
Pamukkale University School of Medicine, Department of Physiology, Denizli, Turkey.
Vural
V
Küçükatay
Ankara University School of Medicine, Department of Forensic Medicine, Ankara, Turkey.
N
N
Lale Şatıroğlu-Tufan
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism*; D009857:Oncogenes / Q000187:drug effects* / Q000502:physiology; D013447:Sulfites / Q000494:pharmacology* / Q000627:therapeutic use; D016896:Treatment Outcome
D009363:Neoplasm Proteins; D013447:Sulfites; C470990:URGCP protein, human
Neuroblastoma; SH-SY5Y cells; Sulfite; URG4/URGCP