Zhang Yan, Zhao Wenjin, Zhao Yujie, He Qun
Department of Biochemistry, College of Basic Medicine, Shenyang Medical College, Shenyang, 110034, Liaoning, China.
Glycoconj J. 2015 Feb;32(1-2):39-47. doi: 10.1007/s10719-014-9569-z. Epub 2015 Jan 9.
We measured ST3Gal, ST6Gal, ST6GalNAc and ST8Sia expression in human hepatic carcinoma cell lines, HepG-2 and SMMC-7721 and normal hepatic cell line, L-02 to reveal the relationship between hepatic carcinoma cell lines sialyltransferases expression and cell membrane sialic acid sugar chains. Membrane sialic acid sugar chains in L-02, HepG-2 and SMMC-7721 cell lines were measured with lectin microarrays to find expression profiles. Expression of 20 sialyltransferases was measured with DNA microarray. qRT-PCR and Western blot were used to verify DNA microarrays data. Siaα 2-3Galβ1-3[Siaα2-6GalNAc]α-R and Siaα 2-6Gal/GalNAc sugar chains in hepatic carcinoma cell lines, HepG-2 and SMMC-7721 were upregulated, and 7differentially expressed sialyltransferases were captured. ST3Gal-IV and ST6Gal I were overexpressed and ST3Gal-I, ST3Gal-V, ST3Gal-VI, ST6GalNAcII and ST6GalNAcVI were downregulated in HepG-2 and SMMC-7721 cell Lines, compared with control cell line. ST6GalNAc-IV and ST8sia expressions were not detected. Other sialyltransferases were not different among cell lines. Results from qRT-PCR and Western blot were consistent with DNA microarray. Overexpression of ST3Gal-IV and ST6Gal I in HepG-2 and SMMC-7721 cell lines may correlate with upregulation of Siaα 2-3Galβ1-3[Siaα2-6GalNAc]α-R and Siaα 2-6Gal/GalNAc sugar chains on cell membranes.
我们检测了人肝癌细胞系HepG-2和SMMC-7721以及正常肝细胞系L-02中ST3Gal、ST6Gal、ST6GalNAc和ST8Sia的表达,以揭示肝癌细胞系唾液酸转移酶表达与细胞膜唾液酸糖链之间的关系。用凝集素微阵列检测L-02、HepG-2和SMMC-7721细胞系中的膜唾液酸糖链,以发现表达谱。用DNA微阵列检测20种唾液酸转移酶的表达。采用qRT-PCR和Western blot验证DNA微阵列数据。肝癌细胞系HepG-2和SMMC-7721中的Siaα 2-3Galβ1-3[Siaα2-6GalNAc]α-R和Siaα 2-6Gal/GalNAc糖链上调,并捕获了7种差异表达的唾液酸转移酶。与对照细胞系相比,HepG-2和SMMC-7721细胞系中ST3Gal-IV和ST6Gal I过表达,而ST3Gal-I、ST3Gal-V、ST3Gal-VI、ST6GalNAcII和ST6GalNAcVI下调。未检测到ST6GalNAc-IV和ST8sia的表达。其他唾液酸转移酶在各细胞系之间无差异。qRT-PCR和Western blot的结果与DNA微阵列一致。HepG-2和SMMC-7721细胞系中ST3Gal-IV和ST6Gal I的过表达可能与细胞膜上Siaα 2-3Galβ1-3[Siaα2-6GalNAc]α-R和Siaα 2-6Gal/GalNAc糖链的上调有关。