Evaluation of Transforming Growth Factor- beta 2, Smad2, and Smad4 Gene Expression in Human Breast Cancer Cell Line (MCF-7) Treated with Aqueous Extract of Trachyspermum copticum (L.) Link Seeds

Background & Aim: Uncontrolled changes in production of TGF-beta and defects in cell signaling pathway molecules, including Smad2 and Smad4, exert a key role in cancer. In our previous study, we showed that treatment of human ovarian cancer cisplatin-resistant and –sensitive cell lines with aqueous extract of Trachyspermum Copticum (L.) Link seeds is associated with reduction in transforming growth factor-beta 2 (TGF-beta 2) production. In this study, we investigated TGF-beta 2, Smad2, and Smad4 gene expression in a human breast cancer cell line (MCF-7) treated with aqueous extract of Trachyspermum copticum (L.) Link seeds. Methods: Non-cytotoxic concentrations of the extract determined by MTT test. MCF-7 cell line treated with non-cytotoxic concentrations (10, 40, and 100 μg/mL) of extract for 24 hours. Then, we did real-time PCR for expression of TGFbeta 2, Smad2, and Smad4 genes. Data were analyzed using REST software. Level of significance was set at 0.05. Results: Data showed a meaningful decrease in expression of TGF-beta 2, and increase in expression of Smad2 and Smad4 genes in cells treated with concentrations of 10 and 40 μg/mL (p<0.0001); while there were no meaningful change in expression of this genes with concentrations of 100 μg/mL (p≥0.05). Conclusion: Results suggest that aqueous extract of Trachyspermum Copticum (L.) Link seeds can affect the TGF-beta 2, Smad2, and Smad4 gene expressions in a time- and dose-dependent manner. Also the extract showed its inhibitory function at low concentrations. The meaningful increase in expression of Smad2 and sSad4 genes in cells treated 24 hours with concentrations of 10 and 40 μg/mL suggests that this extract may be an appropriate tool for modulating the expression of Smad2 and Smad4 gens and thereby, controlling growth and proliferation of cancerous cells. Although further studies, including investigation of simultaneous expression of genes and proteins of TGF-beta 2, Smad2, and Smad4, are required to provide evidences of the effectiveness of Trachyspermum Copticum (L.) Link seeds in vivo.

Keywords:

Trachyspermum copticum (L.) Link , Zenian , Anticancer , Transforming Growth Factor-beta 2 (TGF-beta2) , Smad2 , Smad4 , Cytotoxicity , Real-Time Polymerase Chain Reaction , Real-Time PCR , Human Breast Cancer (MCF7) , Aqueous Extract , Gene Expression ,