Investigation the effect of Thymoquinone on Sox۲ expression in a human fibroblast cell line under cobalt II chloride-induced hypoxic condition

Background and Aim: Fibroblasts are spindle-shaped cells which usually found in the interstitial spaces of organs also they are exciting in a dense environment of cells closing endothelium in the lung. They are the main source of extracellular matrix proteins, in addition, they play crucial roles in determining cell phenotype and function as a result of providing a scaffold for cells. No wonder, fibroblasts contribute to injury responses and stuff like that. Thymoquinone (TQ) is a phytochemical compound found in the plant Nigella sativa with anti-inflammatory and antioxidant effects. In this study, we investigated Sox۲ gene expression in a human fibroblast cell line under hypoxic conditions.Methods: Human fibroblast cells were cultured in DMEM-F۱۲ medium supplemented with ۱۰% fetal bovine serum. Then, the cultured cells were treated with CoCl۲ (۱۰۰ μmol/L) as control group and with CoCl۲ and TQ (۵۰۰ ng/ml) as treatment group for ۲۴ h. To study Sox۲ gene expression at transcription level, total RNA was extracted from the cells. After DNase I treatment of the extracted RNAs and cDNA synthesis, the expression of Sox۲ gene was evaluated by quantitative real-time PCR (qRT-PCR) in treated samples compared to the untreated cells.Results: The results showed ۲۲% reducing Sox۲ gene expression in the treatment group in comparison to the control group as well as the microscopic investigation was shown control group had fewer detachment cells also in morphological view, the control groups had a fresher appearance.Conclusion: This study showed that TQ reduces proliferation in human fibroblast cell line. Furthermore, the previous study had proved that TQ has the antitumor role so it is not far from the imagination that it can be considered as an effective anticancer drug in the future.

Keywords:

Fibroblast cell line , Thymoquinone , CoCl۲ , Hypoxia , Sox۲ gene expression