Interferon-induced transmembrane protein 1 (IFITM1) overexpression enhances the aggressive phenotype of SUM149 inflammatory breast cancer cells in a signal transducer and activator of transcription 2 (STAT2)-dependent manner.

Joshua Ogony,Asona Lui,Massimo Cristofanilli,Joan Lewis-Wambi,Hye Joung Choi

doi:10.1186/s13058-016-0683-7

Abstract

BackgroundInflammatory breast cancer (IBC) is a very aggressive and lethal subtype of breast cancer that accounts for about 4 % of all breast cancers diagnosed in the United States. Despite the efforts of several investigators to identify the molecular factors driving the aggressive phenotype of IBC, a great deal is still unknown about the molecular underpinnings of the disease. In the present study, we investigated the role of interferon-induced transmembrane protein 1 (IFITM1), a well-known interferon-stimulated gene (ISG), in promoting the aggressiveness of SUM149 IBC cells.MethodsWestern blot and real-time polymerase chain reaction analyses were performed to assess the protein and messenger RNA (mRNA) levels of IFITM1 and other ISGs in three IBC cell lines: SUM149, MDA-IBC-3, and SUM190. IFITM1 expression and cellular localization were assessed by using immunofluorescence, while the tumorigenic potential was assessed by performing cell migration, invasion, and colony formation assays. Small interfering RNA and short hairpin RNA knockdowns, enzyme-linked immunosorbent assays, and luciferase assays were performed to determine the functional significance of IFITM1 and signal transducers and activators of transcription 1 and 2 (STAT1/2) in SUM149 cells.ResultsWe found that IFITM1 was constitutively overexpressed at the mRNA and protein levels in triple-negative SUM149 IBC cells, but that it was not expressed in SUM190 and MDA-IBC-3 IBC cells, and that suppression of IFITM1 or blockade of the IFNα signaling pathway significantly reduced the aggressive phenotype of SUM149 cells. Additionally, we found that knockdown of STAT2 abolished IFITM1 expression and IFITM1 promoter activity in SUM149 cells and that loss of STAT2 significantly inhibited the ability of SUM149 cells to proliferate, migrate, invade, and form 2-D colonies. Notably, we found that STAT2-mediated activation of IFITM1 was particularly dependent on the chromatin remodeler brahma-related gene 1 (BRG1), which was significantly elevated in SUM149 cells compared with SUM190 and MDA-IBC-3 cells.ConclusionsThese findings indicate that overexpression of IFITM1 enhances the aggressive phenotype of triple-negative SUM149 IBC cells and that this effect is dependent on STAT2/BRG1 interaction. Further studies are necessary to explore the potential of IFITM1 as a novel therapeutic target and prognostic marker for some subtypes of IBCs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-016-0683-7) contains supplementary material, which is available to authorized users.

Highlights

Inflammatory breast cancer (IBC) is a very aggressive and lethal subtype of breast cancer that accounts for about 4 % of all breast cancers diagnosed in the United States
Ogony et al Breast Cancer Research (2016) 18:25 (Continued from previous page). These findings indicate that overexpression of interferon-induced transmembrane protein 1 (IFITM1) enhances the aggressive phenotype of triple-negative SUM149 IBC cells and that this effect is dependent on STAT2/Brahma-related gene 1 (BRG1) interaction
IFITM1 is overexpressed in triple-negative SUM149 inflammatory breast cancer cells To identify the factors contributing to the aggressive phenotype of IBC cells, we measured the expression of IFITM1, an interferon-stimulated gene (ISG) linked to tumor progression, in three IBC cell lines: triple-negative SUM149, human epidermal growth factor receptor 2 (HER2)-amplified SUM190 and MDA-IBC-3, and the non-IBC cell line MCF-7 (ER+)

Summary

Introduction

Inflammatory breast cancer (IBC) is a very aggressive and lethal subtype of breast cancer that accounts for about 4 % of all breast cancers diagnosed in the United States. The most notable alterations that have been reported include lack of estrogen and progesterone receptors (ER−/PR−); overexpression of epidermal growth factor receptor, ERBB2/human epidermal growth factor receptor 2 (HER2), E-cadherin, eIF4GI, chemokines, and chemokine receptors; dysfunction of mucin 1; high proliferation; tumor protein 53 mutations; and elevated angiogenesis [6] Despite these efforts, there is still a great deal that is not known about the biology of IBC or the factors that drive its aggressive phenotype. Interferons (IFNs) are cytokines that affect biological responses through the Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling pathway This pathway involves IFNs, which, acting as ligands, bind their corresponding receptors [interferon (alpha, beta and omega) receptor (IFNAR)/interferon gamma receptor], resulting in the phosphorylation and activation of STAT1 and STAT2 and subsequent transcription of interferonstimulated genes (ISGs), which include STAT1, STAT2, phospholipid scramblase 1 (PLSCR1), interferon-induced transmembrane protein 1 (IFITM1), interferon-inducible protein 27 (IFI27), interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), and many others [8]. Increased expression of IFITM1, a well-known ISG, has been shown to correlate with disease progression, resistance to endocrine therapy and chemotherapy, and worse overall prognosis in patients with gastrointestinal, colorectal, and breast cancers [14, 15]

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