反义寡核苷酸对Daudi细胞α2,6唾液酸的下调作用研究

A Study of Antisense-Oligonucleotides in Down-Regulation of α2,6-Sialylation of Daudi Cells

目的 研究反义寡核苷酸对Burkitt’s淋巴瘤Daudi细胞α2 ,6 唾液酸转移酶 (ST6GalI)的活性和细胞表面α2 ,6 唾液酸水平的下调作用。方法 以修饰后的ST6GalI的反义寡核苷酸处理Daudi细胞 ,以RT PCR检测Daudi细胞ST6GalImRNA的表达 ,荧光定量法测定ST6GalI的活性及以流式细胞仪检测细胞表面α2 ,6 唾液酸的含量。结果 和对照组比较 ,以反义寡核苷酸处理的Daudi细胞ST6GalImRNA的表达下调 ,ST6GalI酶活性下降 ,并导致细胞表面α2 ,6 唾液酸水平降低。结论 反义寡核苷酸可有效降低Daudi细胞表面α2 ,6 唾液酸化水平 ,内源性减少α2 ,6 唾液酸对Daudi细胞CD2

Objective To study the antisense oligonucleotides (antisense ODN) in down regulation of α2,6 sialyltransferase (ST6Gal I) activity and α2,6 sialylation of cell surface of Burkitt's lymphoma cell line Daudi.Methods Daudi cells were treated with modified antisense ODN in vitro, the ST6Gal I mRNA expression was examined by RT PCR, the ST6Gal I enzyme activity was measured with fluorometric sialyltransferase assay and the amount of α2,6 sialylation of cell surface was determined by FACS analysis.Results Antisense ODN treatment led to a down regulation of ST6Gal I mRNA expression and enzyme activity of Daudi cells and resulted in a reduced α2,6 sialylation of cell surface compared with the control group.Conclusion Antisense ODN is of relevance in decreasing the α2,6 sialylation of Daudi cell surface and may endogenously reduce the masking of CD22 by the α2,6 sialylation.