Evaluation of hydroalcoholic extract effects of Ferula assa-foetida on expression change of EMT and CD44-related genes in gastric cancer stem cell

Abstract

BackgroundGastric cancer (GC) with high incidence is a life-threatening malignant tumor in humans. Cancer stem cells (CSCs) due to tumor-initiating capacity, ability of invasion, metastasis, recurrence, and chemoresistance may be utilized for cancer treatment. Ferula assa-foetida as a herbal remedy in ancient Iranian medicine was shown to have various biological activities, including anti-cancer. Epithelial-mesenchymal transition (EMT) markers play an important role in migration, invasion, and metastatic dissemination of CSCs. The gastric CSCs-surface glycoprotein CD44 also responsible for high tumorigenicity, self-renewal, and metastasis through EMT. In this study, we investigated the hydroalcoholic extract effects of F. assa-foetida on the expression of EMT-related genes, anti-apoptotic factor Bcl-2 and CD44 cell-surface molecule in CSCs. MethodsCSCs were cultured in serum-free medium containing EGF and B-27 and exposed to different concentrations of F. assa-foetida hydroalcoholic extract and for different time durations. Cytotoxic effects of the extract compounds were evaluated using MTT assay. After CSCs treatment with IC50 dose of the extract, the expression level of EMT markers vimentin, Snail1, and Zeb1 and anti-apoptotic marker Bcl-2 was determined using relative quantitative real-time PCR as well as marker CD44 and CD54 were determined by flow cytometry. ResultsOur MTT assay results demonstrated that cytotoxicity effect of extract compounds was significantly dependent on time and concentration. After exposing CSCs to IC50 doses of extract in different times, mRNA expression levels of EMT markers (vimentin, Snail1, Zeb1) and anti-apoptotic marker Bcl-2, as well as protein expression of stemness marker CD44 and CD54 significantly decreased in treated CSCs compared with untreated (P < 0.05). ConclusionOur data demonstrated that extract compounds of F. assa-foetida were able to decrease EMT markers, Bcl-2 anti-apoptotic factor, surface marker of CD44 and CD54, leading to inhibition of CSCs proliferation as well as killing of them. Therefore, isolation and purification of F. assa-foetida hydroalcoholic extract could be used as a potential agent for cancer treatment.