The challenge of determining the impact of FUT3 tumor-associated polymorphism rs2306969 (-6951 C> T) in invasive breast cancer cells
FUT3 gene is responsible for encode an homonymous α1,3/4-fucosyltransferase involved in the synthesis of sialyl-Lewis antigens. FUT3-fucosylated glycoconjugates play key roles in pathways involved in tumor biology and metastasis, such as cellular ligation to E-selectins, TGF-β-induced epithelial-mesenchymal transition, NK cell-mediated tumor cytotoxicity and apoptosis. Tumor-associated FUT3 promoter polymorphism rs2306969 (-6951 C> T, position related to the gene’s translation start site) has been linked to breast, ovarian and intestinal gastric cancer. Although non-coding polymorphisms accounts for the majority of variations founded in breast cancer, their functional roles are still poorly understood. This study aimed to investigate the impact of different alleles for this variation in FUT3 expression of invasive breast tumors. A luciferase reporter assay was performed using two breast tumor cell lines to evaluate respectively the impact of FUT3 rs2306969 (-6951 CC) and (-6951 TT) on protein expression. Gene and protein expressions were also measured in twenty-nine fresh biopsies of invasive breast tumors. Rs2306969 did not significantly influence FUT3 expression in both used systems. However, this study is defiant since the biological role of this polymorphism in breast cancer and other tumor types could be linked to cis/trans modulation of other genes, respond to different environmental stimuli or impact gene expression only in association with other variations. Rs2306969 did not modulate FUT3 expression in breast tumors under non-stimulated conditions. Nevertheless, our study contributes to the notably challenging task that is to understand how non-coding polymorphisms can drive the overall risk in cancer development.
Keywordsα1,3/4-Fucosyltransferase Breast cancer Non-coding single nucleotide polymorphism SNP functional analysis
We gratefully acknowledge Professor Dayane Gomes (Universidade Federal de Pernambuco) for the support during the execution of the Western blotting assays of this work and Professors João Ricardo Mendes and Sergio Crovella (Universidade Federal de Pernambuco) for provided us the Lipofectamine 3000 reagent and the pGL4 vectors, respectively. This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures performed in this study involving human participants were in accordance with international ethical standards and were approved by the Comité de Ética em Pesquisa Envolvendo Seres Humanos do Centro de Ciências da Saúde da Universidade Federal de Pernambuco (CAAE 06586612.9.0000.5208—No. 140.876).
Written informed consent was obtained from all participants included in this study.
- 5.He M, Wu C, Xu J et al (2014) A genome wide association study of genetic loci that influence tumour biomarkers cancer antigen 19-9, carcinoembryonic antigen and alpha fetoprotein and their associations with cancer risk. Gut 63(1):143–151. https://doi.org/10.1136/gutjnl-2012-303434 CrossRefGoogle Scholar
- 6.Shokeen Y, Sharma NR, Vats A, Taneja V, Minhas S, Jauhri M, Sankaran S, Aggarwal S (2018) Identification of prognostic and susceptibility markers in chronic myeloid leukemia using next generation sequencing. Ethiop J Health Sci 28(2):135–146. https://doi.org/10.4314/ejhs.v28i2.5 CrossRefGoogle Scholar
- 13.Breiman A, Robles MDL, Trécesson SC, Echasserieau K, Bernardeau K, Drickamer K, Imberty A, Barillé-Nion S, Altare F, LePendu J (2016) Carcinoma-associated fucosylated antigens are markers of the epithelial state and can contribute to cell adhesion through CLEC17A (Prolectin). Oncotarget 7(12):14064–14082. https://doi.org/10.18632/oncotarget.7476 CrossRefGoogle Scholar
- 16.Liu LL, Zhao H, Ma TF, Ge F, Chen C, Zhang Y (2015) Identification of valid reference genes for the normalization of RT-qPCR expression studies in human breast cancer cell lines treated with and without transient transfection. PLoS ONE 10(1):e0117058. https://doi.org/10.1371/journal.pone.0117058 CrossRefGoogle Scholar