Expression Assay of PTEN, CDK4, and mTOR Genes and Cell Proliferation in Breast Cancer Cell Line (T47D) under Grape Seed Extract Treatment

Abstract

Background: Breast cancer is a highly heterogeneous disease and the most common malignancy in women. Phosphatase and tensin homolog (PTEN) is a tumor suppressor gene that reduces in breast cancer. This gene is a phosphatase, and by its lipid phosphatase capacity separates a phosphate from PIP3, and produces PIP2. So, it tends to inhibition of phosphatidylinositol-3-kinase/AKT (PI3K/AKT) pathway and a break in its cellular cycle in phase G1, increase in apoptosis, and decrease in proliferation of cancer cells. The mammalian target of rapamycin (mTOR) gene is activated by phospho/AKT, and its expression increases in many cancers. The cyclin-dependent kinase 4 (CDK4) gene also acts as an oncogene and its expression increases in various types of human cancers, including breast cancer. Grape seed extract (GSE) is a reach source of natural antioxidants, and treatment with it can tend to inhibition of cell cycle and finally, inhibition of cancer progression. This study aimed to assess the expression of PTEN, CDK4, and mTOR genes and cell proliferation in breast cancer cell line (T47D) under GSE treatment. Methods: T47D cells were purchased and cultured and treated with GSE of a concentration of 60 μg/ml. MTT and real time-polymerase chain reaction (PCR) methods were used to evaluate cell viability and gene expression. Colony survival assay method was used to assess colony formation. Findings: The amount of cell life in the treatment group was significantly lower than the control group. Moreover, the results of real time PCR showed that the relative expression of PTEN gene was significantly higher than the control group, and also the expression of mTOR and CDK4 genes reduced. Conclusion: The use of GSE can significantly increase the expression of PTEN tumor suppressor gene, and ultimately cause the death of T47D cancer cells. © 2022 Isfahan University of Medical Sciences(IUMS). All rights reserved.