Department of Molecular Biology and Genetics, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, 050040, Kazakhstan.
Appl Biochem Biotechnol. 2013 Oct;171(4):963-74. doi: 10.1007/s12010-013-0409-6. Epub 2013 Aug 4.
The terminal monosaccharide of glycoconjugates on a eukaryotic cell surface is typically a sialic acid (Neu5Ac). Increased sialylation usually indicates progression and poor prognosis of most carcinomas. Here, we utilize two human mammary epithelial cell lines, HB4A (breast normal cells) and T47D (breast cancer cells), as a model system to demonstrate differential surface glycans when treated with sialic acid under nutrient deprivation. Under a starved condition, sialic acid treatment of both cells resulted in increased activities of α2→3/6 sialyltransferases as demonstrated by solid phase assay using lectin binding. However, a very strong Maackia amurensis agglutinin I (MAL-I) staining on the membrane of sialic acid-treated T47D cells was observed, indicating an increase of Neu5Acα2→3Gal on the cell surface. To our knowledge, this is a first report showing the utility of lectins, particularly MAL-I, as a means to discriminate between normal and cancer cells after sialic acid treatment under nutrient deprivation. This method is sensitive and allows selective detection of glycan sialylation on a cancer cell surface.
糖缀合物在真核细胞表面的末端单糖通常是唾液酸(Neu5Ac)。唾液酸化程度的增加通常表明大多数癌的进展和预后不良。在这里,我们利用两种人乳腺上皮细胞系 HB4A(乳腺正常细胞)和 T47D(乳腺癌细胞)作为模型系统,在营养剥夺下用唾液酸处理时,证明了细胞表面糖的差异。在饥饿状态下,用唾液酸处理两种细胞都导致α2→3/6 唾液酸转移酶活性增加,这可以通过用凝集素结合的固相测定来证明。然而,在经过唾液酸处理的 T47D 细胞的膜上观察到非常强烈的美洲商陆凝集素 I(MAL-I)染色,表明细胞表面 Neu5Acα2→3Gal 的增加。据我们所知,这是第一个报告,表明在营养剥夺下用唾液酸处理后,凝集素(特别是 MAL-I)可用作区分正常细胞和癌细胞的一种手段。该方法灵敏,可选择性检测癌细胞表面糖基化的唾液酸化。