Bioinformatics and Experimental Evaluation of UBE2W and SSX2IP Expression Levels in HepG2 Cancer Cells Treated with Melatonin

Background and purpose: The aim of this study was to bioinformatically and experimentally evaluate the effect of melatonin on the expression levels of UBE2W and SSX2IP genes in melatonin treated Human Hepatocellular carcinoma G2 (HepG2) cancer cell line. UBE2W encodes a protein that promotes ubiquitination of Fanconi anemia complementation group proteins and may be important in the repair of DNA damage. SSX2IP belongs to an adhesion system, involved in cell movement and acts as a centrosome maturation factor. Materials and methods: At first, as a bioinformatics tool, MATLAB 2018a software was employed to evaluate the UBE2W and SSX2IP genes expression levels using microarray data. Then, after designing and preparing the primers, MTT and Real-time PCR were carried out in three replicates. Results: MTT assay showed that the viability of cancer cell significantly decreased in melatonin treated group (P≤0.01). The IC50 value was estimated to be 2080 µM for 48 hours, but no significant changes were seen in the survival of human normal cells (HUVEC-C) (P≥0.05). Real-time PCR proved that the expression levels of both genes increased (P≥0.05 in 24h and P≤ 0.05 in 48h) in melatonin treated cells compared to un-treated group, which was in accordance with bioinformatics analysis. Conclusion: Our study showed that UBE2W and SSX2IP genes could be used as therapeutic target genes for Hepatocellular carcinoma. However, complementary studies are needed to prove current findings.