Effect of Silibinin on Maspin and ERα Gene Expression in MCF-7 Human Breast Cancer Cell Line

Ghorban Mohammadzadeh,Hossein Babaahmadi-Rezaei,Maryam Karimi,Mohammad-Ali Ghaffari

doi:10.30699/ijp.2017.24871

Ghorban Mohammadzadeh, Hossein Babaahmadi-Rezaei + Show 2 more

Open Access

https://doi.org/10.30699/ijp.2017.24871

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Abstract

According to reports, a serine protease inhibitor (Maspin) suppresses metastasis, invasion and angiogenesis in breast and prostate cancers. Silibinin is a natural polyphenolic flavonoid with anti-cancer activity. We assessed the effects of silibinin on cell viability, maspin and ERα gene expression in MCF-7 cell line. The human MCF-7 breast cancer cell line was cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with different concentrations of silibinin (100-600 μg/mL) for 24, 48 and 72 hours. The cytotoxic effect of silibinin on MCF-7 viability was determined using Methyl-Thiazolyl-Tetrazolium (MTT) assay by IC50 determination. The fold changes of Maspin and ERα expression were determined by reverse-transcription real-time Polymerase Chain Reaction (PCR). All experiments on the cells were performed in triplicates. The maximum inhibitory effect of silibinin on cell viability was observed at 600 μg/mL after 72-hour incubation (p = 0.001). Incubation of the cells with silibinin for 48 and 72 hours significantly decreased IC50 values to 250 and 207 μg/mL (p = 0.005 and p= 0.006), respectively. The expression of maspin and ERα in the treated cells compared to controls was significantly decreased following treatment with different concentrations of silibinin during a 24-hour period. Silibinin reduces both maspin and ERα gene expression in MCF-7 cell line. The therapeutic effect of silibinin on the treatment of breast cancer may be mediated by the reduction of ERα expression. For verifying this hypothesis and the possible therapeutic implication of silibinin on breast cancer, further studies in this direction are necessary.

Highlights

Maspin, a relatively unique member of the serine protease inhibitor family, was originally identified from normal mammary epithelial cells by hybridization assay, according to its expression at the mRNA level [1, 2]
The medium of all wells was removed carefully, and 20 μL of MTT (1 mg/mL dissolved in Phosphate Buffer Saline (PBS)) and 180 μL of Dulbecco's modified Eagle's medium (DMEM) was added to each well and kept for 4.5 hours in the dark for further incubation
The growth of the MCF-7 cell line was inhibited by silibinin in a dose- and time-dependent manner

Summary

Introduction

Maspin (mammary serpin), a relatively unique member of the serine protease inhibitor family, was originally identified from normal mammary epithelial cells by hybridization assay, according to its expression at the mRNA level [1, 2]. Maspin induces apoptosis, and inhibits progression of cancer cell, angiogenesis and invasion, both in cell culture and animal models [3,4,5]. The Maspin gene expression in human breast cancer is paradoxical, since, underand overexpression have been reported [6, 7]. A serine protease inhibitor (Maspin) suppresses metastasis, invasion and angiogenesis in breast and prostate cancers. We assessed the effects of silibinin on cell viability, maspin and ERα gene expression in MCF-7 cell line.

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