Laboratory of Biochemistry, School of Medicine, University of Crete, 70013 Heraklion, Greece.
Institute of Molecular Biology & Biotechnology-FoRTH, Heraklion, 70013 Heraklion, Greece.
Int J Mol Sci. 2021 Jul 29;22(15):8165. doi: 10.3390/ijms22158165.
In breast cancer, expression of Cluster of Differentiation 24 (CD24), a small GPI-anchored glycoprotein at the cell periphery, is associated with metastasis and immune escape, while its absence is associated with tumor-initiating capacity. Since the mechanism of CD24 sorting is unknown, we investigated the role of glycosylation in the subcellular localization of CD24. Expression and localization of wild type N36- and/or N52-mutated CD24 were analyzed using immunofluorescence in luminal (MCF-7) and basal B (MDA-MB-231 and Hs578T) breast cancer cells lines, as well as HEK293T cells. Endogenous and exogenously expressed wild type and mutated CD24 were found localized at the plasma membrane and the cytoplasm, but not the nucleoplasm. The cell lines showed different kinetics for the sorting of CD24 through the secretory/endocytic pathway. N-glycosylation, especially at N52, and its processing in the Golgi were critical for the sorting and expression of CD24 at the plasma membrane of HEK293T and basal B type cells, but not of MCF-7 cells. In conclusion, our study highlights the contribution of N-glycosylation for the subcellular localization of CD24. Aberrant N-glycosylation at N52 of CD24 could account for the lack of CD24 expression at the cell surface of basal B breast cancer cells.
在乳腺癌中,细胞外周的小 GPI 锚定糖蛋白簇分化抗原 24(CD24)的表达与转移和免疫逃避有关,而其缺失与肿瘤起始能力有关。由于 CD24 分拣的机制尚不清楚,我们研究了糖基化在 CD24 亚细胞定位中的作用。使用免疫荧光法在腔(MCF-7)和基底 B(MDA-MB-231 和 Hs578T)乳腺癌细胞系以及 HEK293T 细胞中分析野生型 N36-和/或 N52 突变 CD24 的表达和定位。内源性和外源性表达的野生型和突变型 CD24 定位于质膜和细胞质,但不在核质中。这些细胞系显示出通过分泌/内吞途径分拣 CD24 的不同动力学。N-糖基化,特别是在 N52 处,以及其在高尔基体中的加工,对于 HEK293T 和基底 B 型细胞的质膜上 CD24 的分拣和表达至关重要,但对于 MCF-7 细胞则不重要。总之,我们的研究强调了 N-糖基化对 CD24 亚细胞定位的贡献。CD24 中 N52 处的异常 N-糖基化可能导致基底 B 型乳腺癌细胞表面缺乏 CD24 表达。