Abstract
Background
A specific sialyl-transferases called ST6GALNAC1 has been proven to up-regulate abnormal O‐glycosylation, which is strongly associated with tumorigenesis and cancer progression. However, the precise pathological outcome of ST6GALNAC1 expression in breast cancer cells remains unknown. Therefore, our study aims to investigate the functional role of ST6GALNAC1 and its impact on the epithelial-mesenchymal transition (EMT) pathway in breast cancer cells.
Methods
Plasmids with siRNA were used to construct ST6GALNAC1 knockoff (si-ST6GALNAC1) MDA-MB-231 and MDA-MB-453 cells, while lentiviruses were used to construct ST6GALNAC1 over-expression (oe-ST6GALNAC1) MCF-7 and BT474 cells. Transfer efficiency was verified by Western Blot. Then we selected transfected cells and assessed the changes in cell proliferation, invasion, migration, and EMT markers.
Results
The expression of ST6GALNAC1 significantly enhanced cell migration and invasion, which was confirmed by Wound Scratch Assay and Transwell Assay. Particularly, ST6GALNAC1 expression directly induced the EMT signaling pathway. E-cadherin was markedly decreased in oe-ST6GALNAC1 cells, accompanied by an up-regulation of mesenchymal markers including N-cadherin, snail, and ZEB1. However, no significant correlation was found between ST6GALNAC1 expression and cell proliferation. All of the outcomes were reversely validated in si-ST6GALNAC1 cells.
Conclusions
The expression of ST6GALNAC1 promotes cell migration and invasion probably by triggering the molecular process of the EMT pathway in breast cancer cells, which may provide new clues for designing novel molecular targeted drugs in breast cancer treatment.
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Data availability
Data available within the article or its supplementary materials.
References
An Y, Han W, Chen X, Zhao X, Lu D, Feng J, Yang D, Song L, Yan X (2013) A novel anti-sTn monoclonal antibody 3P9 Inhibits human xenografted colorectal carcinomas. J Immunother 36:20–28. https://doi.org/10.1097/CJI.0b013e31827810d1
Andergassen U, Liesche F, Kölbl AC, Ilmer M, Hutter S, Friese K, Jeschke U (2015) Glycosyltransferases as markers for early tumorigenesis. Biomed Res Int 2015:792672. https://doi.org/10.1155/2015/792672
Carrascal MA, Severino PF, Guadalupe Cabral M, Silva M, Ferreira JA, Calais F, Quinto H, Pen C, Ligeiro D, Santos LL et al (2014) Sialyl Tn-expressing bladder cancer cells induce a tolerogenic phenotype in innate and adaptive immune cells. Mol Oncol 8:753–765. https://doi.org/10.1016/j.molonc.2014.02.008
Dong X, Jiang Y, Liu J, Liu Z, Gao T, An G, Wen T (2018) T-synthase deficiency enhances oncogenic features in human colorectal cancer cells via activation of epithelial-mesenchymal transition. Biomed Res Int 2018:9532389. https://doi.org/10.1155/2018/9532389
Freire-de-Lima L, Gelfenbeyn K, Ding Y, Mandel U, Clausen H, Handa K, Hakomori SI (2011) Involvement of O-glycosylation defining oncofetal fibronectin in epithelial-mesenchymal transition process. Proc Natl Acad Sci U S A 108:17690–17695. https://doi.org/10.1073/pnas.1115191108
Fu C, Zhao H, Wang Y, Cai H, Xiao Y, Zeng Y, Chen H (2016) Tumor-associated antigens: Tn antigen, sTn antigen, and T antigen. Hla 88:275–286. https://doi.org/10.1111/tan.12900
Garbar C, Mascaux C, Merrouche Y, Bensussan A (2018) Triple-negative and HER2-overexpressing breast cancer cell sialylation impacts tumor microenvironment T-lymphocyte subset recruitment: a possible mechanism of tumor escape. Cancer Manag Res 10:1051–1059. https://doi.org/10.2147/CMAR.S162932
Harbeck N, Gnant M (2017) Breast cancer. Lancet 389:1134–1150. https://doi.org/10.1016/s0140-6736(16)31891-8
Hong R, Xu B (2022) Breast cancer: an up-to-date review and future perspectives. Cancer Commun (lond) 42:913–936. https://doi.org/10.1002/cac2.12358
Jin Y, Zhang Y, Li B, Zhang J, Dong Z, Hu X, Wan Y (2019) TRIM21 mediates ubiquitination of snail and modulates epithelial to mesenchymal transition in breast cancer cells. Int J Biol Macromol 124:846–853. https://doi.org/10.1016/j.ijbiomac.2018.11.269
Julien S, Adriaenssens E, Ottenberg K, Furlan A, Courtand G, Vercoutter-Edouart AS, Hanisch FG, Delannoy P, Le Bourhis X (2006) ST6GalNAc I expression in MDA-MB-231 breast cancer cells greatly modifies their O-glycosylation pattern and enhances their tumourigenicity. Glycobiology 16:54–64. https://doi.org/10.1093/glycob/cwj033
Kvorjak M, Ahmed Y, Miller ML, Sriram R, Coronnello C, Hashash JG, Hartman DJ, Telmer CA, Miskov-Zivanov N, Finn OJ et al (2020) Cross-talk between colon cells and macrophages increases ST6GALNAC1 and MUC1-sTn expression in ulcerative colitis and colitis-associated colon cancer. Cancer Immunol Res 8:167–178. https://doi.org/10.1158/2326-6066.Cir-19-0514
Li J, Jiang Z (2022) Chinese society of clinical oncology breast cancer (CSCO BC) guidelines in 2022: stratification and classification. Cancer Biol Med 19:769–773. https://doi.org/10.20892/j.issn.2095-3941.2022.0277
Li L, Lee KM, Han W, Choi JY, Lee JY, Kang GH, Park SK, Noh DY, Yoo KY, Kang D (2010) Estrogen and progesterone receptor status affect genome-wide DNA methylation profile in breast cancer. Hum Mol Genet 19:4273–4277. https://doi.org/10.1093/hmg/ddq351
Liu Z, Liu J, Dong X, Hu X, Jiang Y, Li L, Du T, Yang L, Wen T, An G et al (2019) Tn antigen promotes human colorectal cancer metastasis via H-Ras mediated epithelial-mesenchymal transition activation. J Cell Mol Med 23:2083–2092. https://doi.org/10.1111/jcmm.14117
Munkley J (2016) The role of Sialyl-Tn in cancer. Int J Mol Sci 17:275. https://doi.org/10.3390/ijms17030275
Munkley J, Oltean S, Vodák D, Wilson BT, Livermore KE, Zhou Y, Star E, Floros VI, Johannessen B, Knight B et al (2015) The androgen receptor controls expression of the cancer-associated sTn antigen and cell adhesion through induction of ST6GalNAc1 in prostate cancer. Oncotarget 6:34358–34374. https://doi.org/10.18632/oncotarget.6024
Nguyen AT, Chia J, Ros M, Hui KM, Saltel F, Bard F (2017) Organelle specific O-glycosylation drives MMP14 activation, tumor growth, and metastasis. Cancer Cell 32:639-653.e636. https://doi.org/10.1016/j.ccell.2017.10.001
Ogawa T, Hirohashi Y, Murai A, Nishidate T, Okita K, Wang L, Ikehara Y, Satoyoshi T, Usui A, Kubo T et al (2017) ST6GALNAC1 plays important roles in enhancing cancer stem phenotypes of colorectal cancer via the Akt pathway. Oncotarget 8:112550–112564. https://doi.org/10.18632/oncotarget.22545
Park S, Koo JS, Kim MS, Park HS, Lee JS, Lee JS, Kim SI, Park BW (2012) Characteristics and outcomes according to molecular subtypes of breast cancer as classified by a panel of four biomarkers using immunohistochemistry. Breast 21:50–57. https://doi.org/10.1016/j.breast.2011.07.008
Pinho SS, Reis CA (2015) Glycosylation in cancer: mechanisms and clinical implications. Nat Rev Cancer 15:540–555. https://doi.org/10.1038/nrc3982
Prasad CP, Chaurasiya SK, Guilmain W, Andersson T (2016) WNT5A signaling impairs breast cancer cell migration and invasion via mechanisms independent of the epithelial-mesenchymal transition. J Exp Clin Cancer Res 35:144. https://doi.org/10.1186/s13046-016-0421-0
Pudova EA, Lukyanova EN, Nyushko KM, Mikhaylenko DS, Zaretsky AR, Snezhkina AV, Savvateeva MV, Kobelyatskaya AA, Melnikova NV, Volchenko NN et al (2019) Differentially expressed genes associated with prognosis in locally advanced lymph node-negative prostate cancer. Front Genet 10:730. https://doi.org/10.3389/fgene.2019.00730
Sewell R, Bäckström M, Dalziel M, Gschmeissner S, Karlsson H, Noll T, Gätgens J, Clausen H, Hansson GC, Burchell J et al (2006) The ST6GalNAc-I sialyltransferase localizes throughout the Golgi and is responsible for the synthesis of the tumor-associated sialyl-Tn O-glycan in human breast cancer. J Biol Chem 281:3586–3594. https://doi.org/10.1074/jbc.M511826200
Soares R, Marinho A, Schmitt F (1996) Expression of sialyl-Tn in breast cancer. Correlation with prognostic parameters. Pathol Res Pract 192:1181–1186. https://doi.org/10.1016/s0344-0338(96)80148-8
Tamura F, Sato Y, Hirakawa M, Yoshida M, Ono M, Osuga T, Okagawa Y, Uemura N, Arihara Y, Murase K et al (2016) RNAi-mediated gene silencing of ST6GalNAc I suppresses the metastatic potential in gastric cancer cells. Gastric Cancer 19:85–97. https://doi.org/10.1007/s10120-014-0454-z
Thomas D, Sagar S, Caffrey T, Grandgenett PM, Radhakrishnan P (2019) Truncated O-glycans promote epithelial-to-mesenchymal transition and stemness properties of pancreatic cancer cells. J Cell Mol Med 23:6885–6896. https://doi.org/10.1111/jcmm.14572
Urh K, Žlajpah M, Zidar N, Boštjančič E (2021) Identification and validation of new cancer stem cell-related genes and their regulatory microRNAs in colorectal cancerogenesis. Biomedicines. https://doi.org/10.3390/biomedicines9020179
Vajaria BN, Patel PS (2017) Glycosylation: a hallmark of cancer? Glycoconj J 34:147–156. https://doi.org/10.1007/s10719-016-9755-2
Wang SL, Li YX, Song YW, Wang WH, Jin J, Liu YP, Liu XF, Yu ZH (2011) Triple-negative or HER2-positive status predicts higher rates of locoregional recurrence in node-positive breast cancer patients after mastectomy. Int J Radiat Oncol Biol Phys 80:1095–1101. https://doi.org/10.1016/j.ijrobp.2010.03.038
Wang L, Liu Y, Wu L, Sun XL (2016) Sialyltransferase inhibition and recent advances. Biochim Biophys Acta 1864:143–153. https://doi.org/10.1016/j.bbapap.2015.07.007
Wang WY, Cao YX, Zhou X, Wei B, Zhan L, Sun SY (2019) Stimulative role of ST6GALNAC1 in proliferation, migration and invasion of ovarian cancer stem cells via the Akt signaling pathway. Cancer Cell Int 19:86. https://doi.org/10.1186/s12935-019-0780-7
Xu W, Yang Z, Lu N (2015) A new role for the PI3K/Akt signaling pathway in the epithelial-mesenchymal transition. Cell Adh Migr 9:317–324. https://doi.org/10.1080/19336918.2015.1016686
Xu X, Yan Q, Wang Y, Dong X (2017) NTN4 is associated with breast cancer metastasis via regulation of EMT-related biomarkers. Oncol Rep 37:449–457. https://doi.org/10.3892/or.2016.5239
Yu X, Wu Q, Wang L, Zhao Y, Zhang Q, Meng Q, Pawan WS (2016) Silencing of ST6GalNAc I suppresses the proliferation, migration and invasion of hepatocarcinoma cells through PI3K/AKT/NF-κB pathway. Tumour Biol 37:12213–12221. https://doi.org/10.1007/s13277-016-5086-y
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Luo, Y., Cao, H., Lei, C. et al. ST6GALNAC1 promotes the invasion and migration of breast cancer cells via the EMT pathway. Genes Genom 45, 1367–1376 (2023). https://doi.org/10.1007/s13258-023-01445-y
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DOI: https://doi.org/10.1007/s13258-023-01445-y