Dall'Olio F, Malagolini N, Guerrini S, Lau J T, Serafini-Cessi F
Department of Experimental Pathology, University of Bologna, Italy.
Glycoconj J. 1996 Feb;13(1):115-21. doi: 10.1007/BF01049687.
We have previously documented a dramatic elevation in the activity of alpha 2,6-sialytransferase towards Gal beta 1,4GlcNAc (EC 2.4.99.1) (alpha 2,6ST) in CaCo-2 cells maintained in culture for several days after confluence to elicit a high degree of enterocytic differentiation phenotype. Northern analysis performed with a probe complementary to a region of human alpha 2,6ST mRNA common to all known transcripts demonstrated that the expression of alpha 2,6ST mRNA in CaCo-2 cells increased with the degree with the degree of differentiation. When probes complementary to 5'-untranslated exons (Y + Z or X) previously identified in transcripts isolated from human placenta and from several human lymphoblastoid cell lines were used, no hybridization signal with mRNA of CaCo-2 cells was found, as reported for the mRNA of hepatoma cell line HepG2 (Wang XC, Vertino A, Eddy RL, Byers MG, Jani-Sait SN, Shows TB, Lau JTY (1993) J Biol Chem 268: 4355-61). These results support the notion that the major alpha 2,6ST transcript of CaCo-2 cells was the hepatoma isoform or a new one, so far unreported. Consistent with the differentiation-dependent increase in alpha 2,6ST-mRNA expression, an elevation of the reactivity with Sambucus nigra agglutinin of differentiated CaCo-2 cell-surface was observed, indicating an enhanced alpha 2,6-sialylation of membrane glycoconjugates.
我们之前曾记录到,在汇合后培养数天以诱导高度肠细胞分化表型的CaCo-2细胞中,α2,6-唾液酸转移酶对Galβ1,4GlcNAc(EC 2.4.99.1)(α2,6ST)的活性显著升高。用与所有已知转录本共有的人类α2,6ST mRNA区域互补的探针进行Northern分析表明,CaCo-2细胞中α2,6ST mRNA的表达随分化程度增加。当使用与先前在从人胎盘和几种人淋巴母细胞系分离的转录本中鉴定出的5'-非翻译外显子(Y + Z或X)互补的探针时,未发现与CaCo-2细胞mRNA的杂交信号,肝癌细胞系HepG2的mRNA也有此报道(Wang XC,Vertino A,Eddy RL,Byers MG,Jani-Sait SN,Shows TB,Lau JTY(1993)J Biol Chem 268:4355 - 61)。这些结果支持这样一种观点,即CaCo-2细胞的主要α2,6ST转录本是肝癌异构体或一种迄今未报道的新转录本。与α2,6ST - mRNA表达的分化依赖性增加一致,观察到分化的CaCo-2细胞表面与黑接骨木凝集素的反应性升高,表明膜糖缀合物的α2,6-唾液酸化增强。