Multidisciplinary Cancer Investigation Original Article January 2019, Volume 3, Issue 1 Bioinformatics-Based Prediction of FUT8 as a Therapeutic Target in Estrogen Receptor-Positive Breast Cancer Fateme Shaabanpour Aghamaleki1, Shirin Farivar1,* 1 Department of Cellullar-Molecular Biology , Faculty of biological Sciences and technologies, Shahid Beheshti University G.C., Tehran, Iran *Corresponding author: Shirin Farivar,Department of Cellullar-Molecular Biology, Faculty of biological Sciences and technologies, Shahid Beheshti University G.C., Tehran, Iran. Tel: +982129902720; Fax: +982122431664; E-mail: [email protected] DOI: 10.30699/acadpub.mci.3.1.25 Abstract Submitted: 20 October 2018 Introduction: Estrogen receptor-positive (ER-positive) breast cancer is a subgroup of Revised: 4 November 2018 breast tumors that is more likely to respond to hormone therapy. ER-positive and ER- Accepted: 28 November 2018 negative breast cancers tend to show different patterns of metastasis because of different e-Published: 1 January 2019 signaling cascade and genes that are activated by estrogen response. Genetic factors can contribute to high rates of metastasis in ER-positive breast cancer. Fucosyltransferase 8 Keywords: (FUT8) is a member of fucosyltransferases family and plays an important role in α-1,6 Fucosyltransferases linkage to the first GlcNAc residue of N-glycans chain. In this study, for the first time, Breast Neoplasms Microarray Analysis we predicted FUT8 by bioinformatics tools as a novel therapeutic target for ER-positive Signal Transduction breast cancer. Methods: Microarray gene expression data of 9 patients with ER+ve and 10 individuals with ER-ve breast cancer was extracted from Geodatasets. Gene expression of two ER+ and ER- patients was compared with logfc and then sorted by their p-values. Moreover, the most related pathway, protein interaction, and function of this gene were identified with GeneCard and DAVID databases. Results: FUT8 was highly expressed in patients with ER+ve breast cancer that may be associated with the metastasis. FUT8 encodes an enzyme that belongs to fucosyltransferases family. The expression of this gene may contribute to the malignancy features of cancer cells and their invasive and metastatic capabilities. Conclusions: Having in mind FUT8 hyperexpression, its function in malignancy, and its pathways, it can be concluded that FUT8 can be used as a therapeutic target in ER+ve breast cancer. © 2019. Multidisciplinary Cancer Investigation INTRODUCTION Breast cancer is one of the most prevalent receptors, progesterone receptors, and HER2), and malignancies in women with 4 major types according triple negative (these tumors are not positive for to the presence of sex hormone receptors on the estrogen receptors, progesterone receptors, and cancer cell. Endocrine receptor-positive (estrogen HER2) tumors are the main categories of breast or progesterone receptors), HER2-positive, triple cancer [1-3]. About 80% of all breast cancers are positive (these tumors are positive for estrogen estrogen receptor-positive (ER-positive) that may Multidiscip Cancer Invest. January 2019, Volume 3, Issue 1 influence the growth of cancer cells in response ER-ve patients that were submitted in September to the estrogen [4-6]. About 65% of breast tumors 2011 and updated in April 2017. are progesterone receptor-positive (PR-positive) Gene Expression Profile Analysis which grow in response to another sex hormone called progesterone [7-9]. These hormones This profile was analyzed; using limma package in R especially estrogen, play an important role in cell program. Limma is an R/Bioconductor package and cycle and tumor progression [4, 10, 11]. Estrogens a powerful differential expression analyzing tool are considered to play a major role in promoting for RNA-sequencing and microarray studies [20, the proliferation of normal and neoplastic breast 21]. Generated unnormalized data were normalized epithelial cells. Three major mechanisms are by robust multiarray averaging (RMA) method. suggested to be involved in estrogen carcinogenic To determine data value and their normalization, effects: 1. Stimulation of cellular proliferation the distribution of the samples was performed. In through estrogens receptor-mediated hormonal this step, genes were compared based on their log activity. Actually, when hormones bind to the cell fold change (logfc) and their level of expression surface receptors, hormone-responsive genes and between ER+ve and ER-ve breast cancer patients. signaling cascades including MAPK signaling They were then sorted into hyperexpressed genes and PI3K signaling pathways, are turned on for in ER+ve patients and hypoexressed genes in ER- DNA synthesis and cell proliferation. 2. Increasing ve breast cancer individuals according to their mutation level through a cytochrome P450-mediated p-value and vice versa to determine specific gene metabolic activation. Cytochrome P-450 catalyzes in ER+ve breast cancer. Moreover, the expression the oxidative metabolism of estrogen and estradiol level of a gene of interest was determined among to a 2-hydroxycatechol estrogen that can induce all 19 samples through the profile graph in NCBI genetic mutations. 3. Induction of aneuploidy Geodatasets. Finally, a gene that was hyperexpressed including loss of 9p11-13 and 4p15.3-16 deletion and had the lowest p-value in ER+ve breast cancer that have been introduced to be tumorigenic in patients was chosen for further analysis. The most several studies [12-14]. In addition, genetic factors related pathway (Gene Ontology) and function of play an important role in breast cancer initiation the gene of interest were identified by the GeneCard and progression. Among genetics element, somatic and DAVID databases for understanding its effect in mutations happen more frequently in breast cells [15]. the pathogenesis of ER+ breast tumor. However, some patients show a germline mutation in BRCA1 and BRCA2 genes. These genes are used GSE32394/GPL11097,selected sample in many prediction tests for breast cancer in patients with a familial background [16-18]. Some of the important genes are expressed only after activating 12 the estrogen receptor and affect both normal and cancer cells. Therefore, identification of molecular 10 keys in breast cancer can result in better decision making about breast cancer treatment. In this article, 8 Microarray gene expression data of breast cancer patients were studied from Geodatabase to identify 6 critical and different genes between ER-positive and ER-negative breast cancer. 4 METHODS 2 Gene Profile Extraction Microarray gene expression data of 9 cases with ER+ve and 10 patients with ER-ve breast cancer GSM801843 GSM801844 GSM801845 GSM801846 GSM801847 GSM801848 GSM801849 GSM801850 GSM801851 GSM801852 GSM801853 GSM801854 GSM801855 GSM801856 GSM801857 GSM801858 GSM801859 GSM801860 GSM801861 (GSE32394) were extracted from NCBI Geodatasets Figure 1: Distribution of Data Value: Distribution Can Be Viewed by [19]. This profile included samples of ER+ve and Box Plot to Indicate Data Value and Normalization 26 Shaabanpour Aghamaleki et al. motifs. and has SH3_9SH3_1SKI NodZ with 575 aa on14q23.3andencodesaprotein gene islocated gene expression profile analysis (Figure 2).compared toER-vebreast cancerpatientsthrough This with 2.2064logfcinER+vebreastcancerpatients analysis (Figure1).FUT8washyperexpressed expression for gene data microarray a valuable and itsboxplot,itwasconcludedthatcanbe According totheanalysisofdistributeddatavalue RESULTS activity throughSTRINGdatabases. its confirm to identified also was proteins other with of thisgene product of theprotein interaction The 3 Figure Figure 2 Figure Sample value Sample value 10 10 : FUT8 Expression Among All Samples Among : FUT8Expression 5 6 7 8 9 : Different Expression Level of FUT8 Was Observed Among ER+andER-veBreastCancer(GSM IndicatesSamples). Observed Was ExpressionLevelofFUT8 : Different 5 6 7 8 9 expression expression GSM801843 GSM801843 GSM801844 GSM801844 va va lu lu e e GSM801845 GSM801845 GSM801846 ER GSM801846 Positive GSM801847 GSM801847 BC GSE GSM801848 GSE GSM801848 32394/N GSM801849 32394/GPL GSM801849 GSM801850 GSM801850 M_ 178155. GSM801851 11097/NM_ are considered as therapeutic targets forcancer. are consideredastherapeutic targets synthesis pathwayandbiosynthesis ofkeratansulfate N-glycan as EGFR,IGFR,andFGFR. Recently, FUT8, exhibitsinanumber ofglycoproteinssuch and metastasis.N-glycan, which is synthesized by pathway playsanimportantroleincancerinvasion databases(Figure4).N-glycansynthesis and DAVID and transcriptionalmisregulationbasedonGeneCard biosynthesisofkeratansulfate, synthesis pathway, Important pathwaysofFUT8includedN-glycan samples. of the all FUT8 washyperexpressedinnearly that (Figure According 3). to this figure, determined it’s samples all in overexpression its recognize to FUT8 expression was identified among 19 samples GSM801851 GSM801852 GSM801852 1_psr GSM801853 17815 GSM801853 1_ GSM801854 a_at/F GSM801854 5.1 _p GSM801855 UT GSM801855 ER sr 8 1_a_at GSM801856 Ne GSM801856 ga ti ve GSM801857 GSM801857 BC GSM801858 GSM801858 GSM801859 GSM801859 GSM801860 GSM801860 27 GSM801861 GSM801861 Multidiscip Cancer Invest. January 2019, Volume 3, Issue 1 A GLYCOSAMINOGLYCAN BIOSYNTHESIS -KERATAN SULFATE N-Glycan biosynthesis S S S CHST1 CHST6 CHST1 FUcα1 6 6 6 [Gal βı — 4GlcNAcβı]0-40 — [3Gal βı — 4GlcNAcβı]10-12— [3Gal βı — 4GlcNAcβı] — 3Gal βı — 4GlcNAcβı — 2Manα1 FUT8 B4GALT4 B3GNT7 B43ALT4 B3GNT2 B4GALT1,2,3 B3GNT2 B4GALT1,2,3