Evaluation of RAC1 Gene Expression Under Exposure Of Plasma Activated Verbascoside in HT-29 Colorectal Cancer Cells

Abstract

Colon cancer is one of the most common and most prevalent Neoplasma in men; adenocarcinomas make up 95 percent of all colon cancer cases. Due to the side effects of conventional treatment methods such as chemotherapy, today, attentions have been attracted to natural compounds that have less side effects and anticancer properties. This study aims to investigate the effect of Verbascoside as a natural compound in inhibiting HT-29 signaling pathway which is an effective route in the metastasis, angiogenesis and cell migration in the human cell line of HT-29 colon cancer; and also investigates the possibility of enhancing and raising the effect of this herbal drug by cold atmospheric pressure plasma jet. A the first step, HT-29 cells were treated at different concentrations of verbascoside 10, 20, 30, 40, 50, 70, 100 μg / ml for 24 hours, then MTT test was used to calculate the inhibitory concentration. The colon cancer cell migration was evaluated by scratch test and HIF-1α gene expression was evaluated by Real Time PCR. The results of the MTT assay showed that verboscoside dose-dependent inhibition of cancer cells. The HT-29 colon is at a concentration of 50 μg / ml. Scratch repair tests showed that verboscoside reduced the migration of cells in a dose-dependent manner, and the results of Real Time PCR showed a decrease in the expression of HIF-1α gene expression. Cancer tumors undergo some changes in oxygen deficiency conditions, including increasing the expression of the HIF-1α gene, which in turn play an important role in activating and enhancing the expression of genes involved in metastasis, angiogenesis and migration. At the second step of this study, verboscoside was primarily treated by cold atmospheric plasma jet then the same procedure has been taken for plasma activated verboscoside. In this study helium jet has been used. The findings of this study showed that verboscosid with inhibitory expression of HIF-1α has a significant anti-metastatic, cellular angiogenesis and migration effect and may be a good option for reducing metastasis and angiogenesis in colon cancer models; it has been also showed that plasma activated verboscosid is more efficacious in all of the above mentioned features.