Study the effect of cisplatin with melatonin on expression of miR-181b, CYLD, CBX-7, BCL2 and p53 in MG-63 osteosarcoma cells

Abstract

Osteosarcoma (OS) is one of the primary malignant cancers that is more common in adolescents and young adults. Diagnostic tests for OS include laboratory tests and radiography, bone scan, CT scan, or MRI. To avoid the risks of X-rays as well as high diagnostic costs, it is necessary to study the effect of medications on OS cell line. Therefore, the aim of this study was to determine the effect of cisplatin with melatonin on the expression of miR-181b and BCL2, CYLD, CBX-7 and p53 genes in MG-63 OS cell line. Methods and Materials: MG-63 OS cell line was cultured in RPMI-640 medium. The cytotoxic effect of cisplatin and melatonin was assessed by MTT assay. Then total RNA was isolated and cDNA was synthesized. The expression level of miR-181b, CYLD, CBX-7, BCL2 by master mix syber green and specific gene primers. Finally, the rate of apoptosis in MG-63 OS cell line was assessed by AnnexinV / PI and flow cytometry. Results: The cytoxicity of melatonin and cisplatin combination was increased on MG-63 cells. After treatment with cisplatin, melatonin and their combination, the expression of BCL2 was significantly reduced and the expression of miRNA-181b, CBX-7, CYLD and p53 was increased. According to the results of flow cytometry, the combination of cisplatin with melatonin increased apoptosis rate in the MG-63 cells.