J Korean Neurosurg Soc.  2015 Mar;57(3):147-151. 10.3340/jkns.2015.57.3.147.

Comparison of the Effects of 13-cis Retinoic Acid and Melatonin on the Viabilities of SH-SY5Y Neuroblastoma Cell Line

• Tosun M
• Soysal Y
• Mas NG
• Karabekir

• KMID: 2191213
• DOI: http://doi.org/10.3340/jkns.2015.57.3.147

Abstract

OBJECTIVE
Neuroblastoma is one of common childhood tumors. Although its mortality is very high, there is no effective treatment yet. The aim of this project is to evaluate cytotoxic effects of melatonin (MLT) an endogen hormone and 13-cis retinoic acid (13-cis-RA) also named as isotretinoin an analogue of vitamin A on neuroblastoma SH-SY5Y cell line.
METHODS
In this study, SH-SY5Y cell line was used. After cell culture, the cells were exposed to different doses of MLT and 13-cis-RA. 24 and 48 hours later. While the viabilities was estimated with MTT cell viability assay test, apoptotic indexes were calculated after staining with TUNEL based apoptosis kit.
RESULTS
It was observed that MLT has very effective cytotoxic potential than 13-cis-RA on neuroblastoma cell line. At the same time, when MLT and 13-cis-RA were combined, this effect was potentiated. On the other hand, it was found that the effect of 13-cis-RA individually on neuroblastoma cells was very slight.
CONCLUSION
We suggest that in the treatment of patient with neuroblastoma, MLT is very effective and also this effect can be augmented by combination with 13-cis-RA.

Keyword

Neuroblastoma; Melatonin; 13-cis retinoic acid; Cancer; Cell viability; Isotretinoin

MeSH Terms

Apoptosis
Cell Culture Techniques
Cell Line*
Cell Survival
Hand
Humans
In Situ Nick-End Labeling
Isotretinoin
Melatonin*
Mortality
Neuroblastoma*
Tretinoin*
Vitamin A
Isotretinoin
Melatonin
Tretinoin
Vitamin A

Figure

• 

  Fig. 1 A: The viability rates in SH-SY5Y neuroblastoma cell line at the end of 24th and 48th hours (%) (MTT viability assay). B: The apoptotic index in SH-SY5Y neuroblastoma cell line at the end of 24th and 48th hours (%) (TUNEL based apoptosis detection). MTT: microculture tetrazolium, DMSO: dimetylsulfoxide, MLT: melatonin, RA: retinoic acid.

• 

  Fig. 2 A: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to 13-cis-RA 1 µM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). B: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to 13-cis-RA 10 µM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). C: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to MLT 2 mM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). D: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to MLT 0.5 mM+13-cis-RA 1 µM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). E: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to MLT 1 mM+13-cis-RA 5 µM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). F: The immunohistochemically stained SH-SY5Y neuroblastoma cells exposure to MLT 2 mM+13-cis-RA 10 µM (TdtTUNEL based apoptosis detection kit, MK500, Takara). The cells green in color are non-apoptotic and brown in color are apoptotic (×200). MLT: melatonin, RA: retinoic acid.

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