β1,4-Galactosyltransferase V Modulates Breast Cancer Stem Cells through Wnt/β-catenin Signaling Pathway
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose Breast cancer stem cells (BCSCs) contribute to the initiation, development, and recurrence of breast carcinomas. β1,4-Galactosyltransferase V (B4GalT5), which catalyzes the addition of galactose to GlcNAcβ1-4Man of N-glycans, is involved in embryogenesis. However, its role in the modulation of BCSCs remains unknown. Materials and Methods The relationship between B4GalT5 and breast cancer stemness was investigated by online clinical databases and immunohistochemistry analysis. Mammosphere formation, fluorescence-activated cell sorting (FACS), and in-vivo assays were used to evaluate B4GalT5 expression in BCSCs and its effect on BCSCs. B4GalT5 regulation of Wnt/β-catenin signaling was examined by immunofluorescence and Ricinus communis agglutinin I pull-down assays. Cell surface biotinylation and FACS assays were performed to assess the association of cell surface B4GalT5 and BCSCs. Results B4GalT5, but not other B4GalTs, was highly correlated with BCSC markers and poor prognosis. B4GalT5 significantly increased the stem cell marker aldehyde dehydrogenase 1A1 (ALDH1A1) and promoted the production of CD44+CD24–/low cells and the formation of mammospheres. Furthermore, B4GalT5 overexpression resulted in dramatic tumor growth in vivo. Mechanistically, B4GalT5 modified and protected Frizzled-1 from degradation via the lysosomal pathway, promoting Wnt/β-catenin signaling which was hyperactivated in BCSCs. B4GalT5, located on the surface of a small subset of breast carcinoma cells, was not responsible for the stemness of BCSCs. Conclusion B4GalT5 modulates the stemness of breast cancer through glycosylation modification to stabilize Frizzled-1 and activate Wnt/β-catenin signaling independent of its cell surface location. Our studies highlight a previously unknown role of B4GalT5 in regulating the stemness of breast cancer and provide a potential drug target for anticancer drug development.
Abstract
Purpose Breast cancer stem cells (BCSCs) contribute to the initiation, development, and recurrence of breast carcinomas. β1,4-Galactosyltransferase V (B4GalT5), which catalyzes the addition of galactose to GlcNAcβ1-4Man of N-glycans, is involved in embryogenesis. However, its role in the modulation of BCSCs remains unknown. Materials and Methods The relationship between B4GalT5 and breast cancer stemness was investigated by online clinical databases and immunohistochemistry analysis. Mammosphere formation, fluorescence-activated cell sorting (FACS), and in-vivo assays were used to evaluate B4GalT5 expression in BCSCs and its effect on BCSCs. B4GalT5 regulation of Wnt/β-catenin signaling was examined by immunofluorescence and Ricinus communis agglutinin I pull-down assays. Cell surface biotinylation and FACS assays were performed to assess the association of cell surface B4GalT5 and BCSCs. Results B4GalT5, but not other B4GalTs, was highly correlated with BCSC markers and poor prognosis. B4GalT5 significantly increased the stem cell marker aldehyde dehydrogenase 1A1 (ALDH1A1) and promoted the production of CD44+CD24–/low cells and the formation of mammospheres. Furthermore, B4GalT5 overexpression resulted in dramatic tumor growth in vivo. Mechanistically, B4GalT5 modified and protected Frizzled-1 from degradation via the lysosomal pathway, promoting Wnt/β-catenin signaling which was hyperactivated in BCSCs. B4GalT5, located on the surface of a small subset of breast carcinoma cells, was not responsible for the stemness of BCSCs. Conclusion B4GalT5 modulates the stemness of breast cancer through glycosylation modification to stabilize Frizzled-1 and activate Wnt/β-catenin signaling independent of its cell surface location. Our studies highlight a previously unknown role of B4GalT5 in regulating the stemness of breast cancer and provide a potential drug target for anticancer drug development.
참고문헌 (30)
- Bray F, "Global cancer statistics 2018 : GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries", CA Cancer J Clin, 68, pp.394~424 (2018)
- Visvader JE, "Cancer stem cells : current status and evolving complexities", Cell Stem Cell, 10, pp.717~728 (2012)
- Ghasemi F, "Therapeutics strategies against cancer stem cell in breast cancer", Int J Biochem Cell Biol, 109, pp.76~81 (2019)
- Fuster MM, "The sweet and sour of cancer : glycans as novel therapeutic targets", Nat Rev Cancer, 5, pp.526~542 (2005)
- Pinho SS, "Glycosylation in cancer : mechanisms and clinical implications", Nat Rev Cancer, 15, pp.540~555 (2015)
- Wassler M, "Pluripotent stem cells", pp.345~372 (2013), InTech
- Ma Y, "The relationship between early embryo development and tumourigenesis", J Cell Mol Med, 14, pp.2697~2701 (2010)
- Gyorffy B, "An online survival analysis tool to rapidly assess the effect of 22, 277 genes on breast cancer prognosis using microarray data of 1, 809 patients", Breast Cancer Res Treat, 123, pp.725~731 (2010)
- Tang Z, "GEPIA : a web server for cancer and normal gene expression profiling and interactive analyses", Nucleic Acids Res, 45, pp.W98~W102 (2017)
- Muller A, "Involvement of chemokine receptors in breast cancer metastasis", Nature, 410, pp.50~56 (2001)
- Robey RW, "Revisiting the role of ABC transporters in multidrug-resistant cancer", Nat Rev Cancer, 18, pp.452~464 (2018)
- Ciccone V, "Stemness marker ALDH1A1 promotes tumor angio-genesis via retinoic acid", HIF-1alpha, J Exp Clin Cancer Res, pp.2018, 37[DOI]
- Li Y, "Inhibition of KLF4 by statins reverses adriamycin-induced metastasis and cancer stemness in osteosarcoma cells", Stem Cell Reports, 8, pp.1617~1629 (2017)
- Yang A, "MYC inhibition depletes cancer stem-like cells in triple-negative breast cancer", Cancer Res, 77, pp.6641~6650 (2017)
- Youakim A, "Localization of the long form of beta-1, 4-galactosyltransferase to the plasma membrane and Golgi complex of 3T3 and F9 cells by immunofluorescence confocal microscopy", Proc Natl Acad Sci U S A, 91, pp.10913~10917 (1994)
- Lim SK, "Wnt signaling promotes breast cancer by blocking ITCH-mediated degradation of YAP", TAZ transcriptional coactivator WBP2, 2016, pp.76, 6278~6289
- Chung J, "Protein arginine methyltransferase 5(PRMT5)promotes survival of lymphoma cells via activation of WNT", beta-catenin and AKT, J Biol Chem, pp.2019, 294[DOI]
- Rada P, "WNT-3A regulates an Axin1", NRF2complex that regulates antioxidant metabolism in hepatocytes, 2015, pp.22, 555~571
- Cooper GM, "The cell: a molecular approach" (2000), Sinauer Associates Inc
- Sola RJ, "Effects of glycosylation on the stability of protein pharmaceuticals", J Pharm Sci, 98, pp.1223~1245 (2009)
- Sasaki N, "beta4GalT-II is a key regulator of glycosylation of the proteins involved in neuronal development", Biochem Biophys Res Commun, 333, pp.131~137 (2005)
- Chen WS, "Tumor beta-1, 4-galactosyltransferase IV overexpression is closely associated with colorectal cancer metastasis and poor prognosis", Clin Cancer Res, 11(24 Pt 1), pp.8615~8622 (2005)
- Yoshihara T, "Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice", PLoS Genet, 14, pp.e1007545~ (2018)
- Talhaoui I, "Identification of key functional residues in the active site of human {beta}1, 4-galactosyltransferase 7 : a major enzyme in the glycosaminoglycan synthesis pathway", J Biol Chem, 285, pp.37342~37358 (2010)
- Katoh M, "Canonical and non-canonical WNT signaling in cancer stem cells and their niches : cellular heterogeneity, omics reprogramming, targeted therapy and tumor plasticity(Review)", Int J Oncol, 51, pp.1357~1369 (2017)
- Wang H, "Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding", J Biol Chem, 290, pp.1654~1663 (2015)
- Li CW, "Glycosylation and stabilization of programmed death ligand-1 suppresses T-cell activity", Nat Commun, 7, pp.12632~ (2016)
- Xu Y, "N-glycosylation at Asn 402 stabilizes N-cadherin and promotes cell-cell adhesion of glioma cells", J Cell Biochem, 118, pp.1423~1431 (2017)
- Yonamine I, "Sphingosine kinases and their metabolites modulate endolysosomal trafficking in photoreceptors", J Cell Biol, 192, pp.557~567 (2011)
- Rodeheffer C, "Targeted mutations in beta1, 4-galactosyltransferase I reveal its multiple cellular functions", Biochim Biophys Acta, 1573, pp.258~270 (2002)