Synergistic Effects of Tamoxifen and Tranilast on VEGF and MMP-9 Regulation in Cultured Human Breast Cancer Cells

Sara Darakhshan,Arezou Rabzia,Mozafar Khazaei,Ali Bidmeshkipour,Ali Ghanbari

doi:10.7314/apjcp.2013.14.11.6869

Sara Darakhshan, Arezou Rabzia + Show 3 more

Open Access

https://doi.org/10.7314/apjcp.2013.14.11.6869

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Abstract

Vascular endothelial growth factor and matrix metalloproteinases are two important factors for angiogenesis associated with breast cancer growth and progression. The present study was aimed to examine the effects of tamoxifen and tranilast drugs singly or in combination on proliferation of breast cancer cells and also to evaluate VEGF and MMP-9 expression and VEGF secretion levels. Human breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with tamoxifen and/or tranilast alone or in combination and percentage cell survival and proliferative activity were evaluated using LDH leakage and MTT assays. mRNA expression and protein levels were examined by real-time RT-PCR and ELISA assay, respectively. LDH and MTT assays showed that the combined treatment of tamoxifen and tranilast resulted in a significant decrease in cell viability and cell proliferation compared with tamoxifen or tranilast treatment alone, with significant decrease in VEGF mRNA and protein levels. We also found that tamoxifen as a single agent rarely increased MMP-9 expression. A decrease in MMP-9 expression was seen after treatment with tranilast alone and in the combined treatment MMP-9 mRNA level was decreased. This combination treatment can able to inhibit growth, proliferation and angiogenesis of breast cancer cells.

Highlights

Materials and Methods: Human breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with tamoxifen and/or tranilast alone or in combination and percentage cell survival and proliferative activity were evaluated using Lactate dehydrogenase (LDH) leakage and MTT assays. mRNA expression and protein levels were examined by real-time RT-PCR and ELISA assay, respectively
LDH and MTT assays showed that the combined treatment of tamoxifen and tranilast resulted in a significant decrease in cell viability and cell proliferation compared with tamoxifen or tranilast treatment alone, with significant decrease in Vascular endothelial growth factor (VEGF) mRNA and protein levels
The results show that the cytotoxic and anti-proliferative effects of tamoxifen and/or tranilast increased in a dose-dependent manner in both MCF-7 and MDA-MB-231 cell lines (Figure 1 and 2)

Summary

Introduction

The formation of new blood capillaries from preexisting blood vessels (Bergers and Benjamin, 2003), supports growth and development of the tumor mass and is an essential process during growth and progression of breast cancer (Banerjee et al, 2007). High VEGF expression increase the micro-vascular density of tumor tissue (Ahluwalia et al, 2012) and has several effects including endothelial cell proliferation, migration, invasion and survival (Banerjee et al, 2007). It have been reported that VEGF stimulates both paracrine and autocrine signaling in endothelial cells, which promotes the progression and survival of tumor cells (Ferrara et al, 2003). It is critical in tumor growth, invasiveness and metastasis of cancer (Folkman, 1990). The effects of this combined treatment on the VEGF mRNA expression and protein level as well MMP-9 mRNA level were examined

Materials and Methods

Results

Discussion