The effect of verbascoside on HIF-1α expression as valuable metastasis and angiogenesis factor in HT-29 colon cancer cells

Colon cancer is one of the most important and most common Neoplasms in men. In 95% of cases of colon cancer is the adenocarcinoma type. Due to the side effects of conventional treatment methods such as chemotherapy, a lot of attention has attracted to natural ingredients that have fewer side effects and anti-cancer properties. The aim of this study is to evaluate the effect of verbascoside as a natural compound in inhibiting HIF-1α signaling pathway, as an effective way of metastasis, angiogenesis and cell migration in human cell line HT-29 colon cancer. HT-29 cell line cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin and streptomycin at 37 ˚C and 5% CO2. HT-29 cells were treated with different concentrations of verbascoside (10,20,30,40,50,70,100 μg/ml) for 24 hours, then MTT assay was used to calculate inhibitory concentration of verbascoside. The migration of the colon cancer cells was evaluated by scratch test and HIF-1α gene expression was evaluated by Real Time PCR. The results of MTT test showed that verbascoside dose-dependently inhibited HT-29 colon cancer cells and IC50 was evaluated as 50 μg/ml. The results of wound healing assay demonstrated Verbascoside decreased cell migration as dose dependent manner. The results of Real Time PCR demonstrated attenuating effect of Verbascoside on HIF-1α gene expression. Cancerous tumors have been altered in hypoxic conditions and induced circumstance such as HIF-1α overexpression which plays an important role in the activation and expression of metastasis, angiogenesis and migration. Our findings showed that verbascoside induced significant anti-metastatic, anti-angiogenic activities and it may be a good candidate to reduce metastasis and angiogenesis in colon cancer studies.

Keywords:

Verbascoside , HIF-1α , metastasis , angiogenesis , HT-29 cell line