Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells
Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells
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DOI：
10.1023/a:1006924208135
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发表时间：
1998-12-01
影响因子：
3
通讯作者：
Hollingsworth, MA
中科院分区：
文献类型：
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作者：
Mack, DR;Cheng, PW;Hollingsworth, MA
为了确定细胞生长条件是否影响碳水化合物表达，将HT 29细胞从常规的含葡萄糖培养基逐渐转移到含无葡萄糖半乳糖的培养基。丙酮固定的细胞上的间接免疫荧光显示，与在含葡萄糖的培养基中生长的那些相比，在无葡萄糖的含半乳糖的培养基中生长的HT 29细胞表面上的唾液酸化刘易斯A抗原（CA 19 -9）、唾液酸化刘易斯C（DUPAN 2）和Tn/唾液酸化-Tn的表达增加。负责合成这些唾液酸化表位的唾液酸转移酶在半乳糖喂养的HT 29细胞中增加。将覆盖细胞的培养基在氯化铯中进行等密度超离心，并使用Mono-Q阴离子交换柱通过HPLC进一步分离衍生自葡萄糖和半乳糖培养基的具有1.56 g/L当量浮力密度的级分，其预测含有粘蛋白糖型。来自在含半乳糖的培养基中生长的细胞的样品的洗脱液的色谱显示与抗唾液酸刘易斯A抗体CA 19 -9反应的峰增加。这些结果表明，体外培养条件的改变可能导致HT 29结肠癌细胞改变唾液酸化碳水化合物的表达。
To determine whether cell growth conditions impacted carbohydrate expression, HT29 cells were gradually transferred from a conventional glucose-containing media to a glucose-free galactose containing media. Indirect immunofluorescence on acetone fixed cells showed increased expression of sialyl Lewis A antigen (CA19-9), sialyl Lewis C (DUPAN2) and Tn/sialyl-Tn on the surface of HT29 cells grown in the glucose-free galactose containing media compared to those grown in the glucose containing media. Sialyltransferases responsible for the synthesis for these sialylated epitopes were increased in the galactose-fed HT29 cells. Media overlying the cells was subjected to isopycnic ultracentrifugation in cesium chloride and the fractions derived from both glucose and galactose media with equivalent buoyant densities of 1.56 g/L, which are predicted to contain mucin glycoforms, were further separated by HPLC using a Mono-Q anion exchange column. The chromatograph of eluent from the sample derived from the cells growing in the galactose containing media showed an increased peak that reacted with the anti-sialyl Lewis A antibody, CA19-9. These results show that alteration of in vitro culture conditions may cause HT29 colonic carcinoma cells to alter the expression of sialylated carbohydrates.