Investigation of anticancer mechanism of oleuropein via cell cycle and apoptotic pathways in SH-SY5Y neuroblastoma cells

• Published in: Gene 2016-07, Vol.585 (1), p.93-99
• Main Authors: Seçme, Mücahit, Eroğlu, Canan, Dodurga, Yavuz, Bağcı, Gülseren
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents - pharmacology; Antioxidants - pharmacology; Apoptosis; Apoptosis - drug effects; Cell cycle; Cell Division - drug effects; Cell Line, Tumor; Cell Movement - drug effects; Cell Proliferation - drug effects; Cell Survival - drug effects; G1 Phase Cell Cycle Checkpoints - drug effects; Gene Expression Regulation, Neoplastic - drug effects; Humans; In Situ Nick-End Labeling; Iridoids - pharmacology; Neoplasm Invasiveness - pathology; Neuroblastoma; Neuroblastoma - drug therapy; Neuroblastoma - pathology; Olea; Oleuropein; Wound Healing - drug effects
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2016.03.038

Neuroblastoma is one of the most common types of pediatric tumors that can spread quickly in neuronal tissues. Oleuropein which is active compound of olive leaves, belongs to polyphenols group and has antioxidant, anti-microbial, anti-inflammatory, anti-hypertensive and anti-carcinogenic effects. The aim of the study is to determine the therapeutic effects of oleuropein on cell proliferation, invasion, colony formation, cell cycle and apoptotic mechanisms in SH-SY5Y neuroblastoma cell line under in vitro conditions. The effect of oleuropein on cell viability was determined by XTT method. 84 cell cycle control and 84 apoptosis related genes were evaluated by RT-PCR. Effects of oleuropein on apoptosis were researched by TUNEL assay. Protein expressions were determined by western blot analysis. Effects of oleuropein on cell invasion, colony formation and migration were detected by matrigel-chamber, colony formation assay and wound-healing assay, respectively. IC50 value of oleuropein in SH-SY5Y cells was detected as 350μM at 48th hours. It is determined that oleuropein causes cell cycle arrest by down-regulating of CylinD1,CylinD2,CyclinD3,CDK4,CDK6 and up-regulating of p53 and CDKN2A, CDKN2B, CDKN1A gene expressions. Oleuropein also induces apoptosis by inhibiting of Bcl-2 and activating of Bax,caspase-9 and caspase-3 gene expressions. Apoptotic cell ratio was found 36.4±3.27% in oleuropein dose group. Oleuropein decreased invasion in SH-SY5Y cells and suppressed colony numbers in ratio of 53.6±4.71%.Our results demonstrated that oleuropein can be a therapeutic agent in the treatment of neuroblastoma. •Oleuropein is inhibiting cell proliferation in SH-SY5Y neuroblastoma cells by inducing cell cycle arrest via genes expression changes of CCND1,CCND2, CDK4, CDK6,p53 and CDK inhibitors.•Oleuropein is stimulating apoptotic mechanisms in SH-SY5Y via intrinsic pathway.•Oleuropein is decreasing invasion, migration and colony formation in SH-SY5Y neuroblastoma cells.