Sulforaphane induces cell cycle arrest by protecting RB-E2F-1 complex in epithelial ovarian cancer cells

Christopher S Bryant,Ramesh B Batchu,Masood A Shammas,Assaad Semaan,Jagannath Pal,Jay Shah,Shelly Seward,Madhu Prasad,Sreedhar Chamala,Mahdi Haider,Sanjeev Kumar,Donald W Weaver,Aamer M Qazi,Robert Morris,Christopher Steffes,Ravindra B Potti

doi:10.1186/1476-4598-9-47

Abstract

BackgroundSulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer. In-vitro exposure to SFN appears to result in the induction of apoptosis and cell-cycle arrest in a variety of tumor types. However, the molecular mechanisms leading to the inhibition of cell cycle progression by SFN are poorly understood in epithelial ovarian cancer cells (EOC). The aim of this study is to understand the signaling mechanisms through which SFN influences the cell growth and proliferation in EOC.ResultsSFN at concentrations of 5 - 20 μM induced a dose-dependent suppression of growth in cell lines MDAH 2774 and SkOV-3 with an IC50 of ~8 μM after a 3 day exposure. Combination treatment with chemotherapeutic agent, paclitaxel, resulted in additive growth suppression. SFN at ~8 μM decreased growth by 40% and 20% on day 1 in MDAH 2774 and SkOV-3, respectively. Cells treated with cytotoxic concentrations of SFN have reduced cell migration and increased apoptotic cell death via an increase in Bak/Bcl-2 ratio and cleavage of procaspase-9 and poly (ADP-ribose)-polymerase (PARP). Gene expression profile analysis of cell cycle regulated proteins demonstrated increased levels of tumor suppressor retinoblastoma protein (RB) and decreased levels of E2F-1 transcription factor. SFN treatment resulted in G1 cell cycle arrest through down modulation of RB phosphorylation and by protecting the RB-E2F-1 complex.ConclusionsSFN induces growth arrest and apoptosis in EOC cells. Inhibition of retinoblastoma (RB) phosphorylation and reduction in levels of free E2F-1 appear to play an important role in EOC growth arrest.

Highlights

Sulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer
Epidemiological observations indicate that cruciferous vegetables such as broccoli, cabbage, cauliflower and brussels sprouts have been shown to offer protection against various cancers [5,6]
SFN induces growth arrest in ovarian cancer cell lines To determine if SFN had potential to induce growth arrest in ovarian cancer cells, we conducted cell proliferation assays using two human ovarian cancer cell lines, MDAH-2774 and SkOV-3

Summary

Introduction

Sulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer. Cytoreductive surgery followed by adjuvant combination chemotherapy is the standard treatment for advanced stage disease Despite these interventions, long-term survival rates remain low underscoring the need for new effective drugs or drug combinations with tolerable side effects [3,4]. Epidemiological observations indicate that cruciferous vegetables such as broccoli, cabbage, cauliflower and brussels sprouts have been shown to offer protection against various cancers [5,6]. Phytochemicals such as glucosinolates are abundant in these vegetables that are converted into isothiocyanates such as sulforaphane (SFN) [6,7,8]

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