α2,6-Hyposialylation of c-Met abolishes cell motility of ST6Gal-I-knockdown HCT116 cells
α2,6-Hyposialylation of c-Met abolishes cell motility of ST6Gal-I-knockdown HCT116 cells
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DOI：
10.1038/aps.2009.84
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发表时间：
2009-07-01
影响因子：
8.2
通讯作者：
Ding, Jian
中科院分区：
文献类型：
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作者：
Qian, Jin;Zhu, Cai-hua;Ding, Jian
目的：研究不同于整合素的α-2，6-唾液酸化修饰先前未知的表面糖蛋白(S)的可能性。方法：通过小干扰RNA降低结肠癌细胞株HCT116中β-半乳糖苷α2，6-唾液酸转移酶(ST6Gal-I)的表达。采用贴壁法和Boyden小室实验检测细胞活力的变化。凝集素亲和分析检测α-2，6-唾液酸化蛋白。结果：在HCT116细胞中，ST6Gal-I基因的敲除抑制了细胞的运动，但不影响细胞的黏附。这种选择性改变的细胞迁移是由于c-Met上α-2，6-唾液酸结构的丢失引起的。在ST6Gal-I-Knokdown(ST6Gal-I-KD)HCT116细胞中，STAT3在酪氨酸705位发生去磷酸化。结论：C-Met是ST6Gal-I的底物。C-Met的低激活可使ST6Gal-I-KD HCT116细胞的运动性丧失。
Aim: We aimed to investigate the potential modification of previously unrecognized surface glycoprotein(s) by alpha 2,6-sialylation other than by integrins.Methods: The expression of beta-galactoside alpha 2,6-sialyltransferase (ST6Gal-I) in the colon cancer cell line HCT116 was reduced by siRNA. The adhesion and Boyden chamber assay were used to detect the variation in cell motility. alpha 2,6-Sialylation proteins were detected with lectin affinity assay. The mRNA expression, protein expression and downstream signaling modulation with siRNA were detected using reverse transcription-polymerase chain reaction, flow cytometry analysis, and Western blot.Results: In HCT116 cells, the knockdown of ST6Gal-I inhibited cell motility, but did not affect cell adhesion. This selectively altered cell migration was caused by the loss of alpha 2,6-sialic acid structures on c-Met. Moreover, STAT3 was dephosphorylated at tyrosine 705 in ST6Gal-I-knockdown (ST6Gal-I-KD) HCT116 cells.Conclusion: c-Met is the substrate of ST6Gal-I. The hyposialylation of c-Met can abolish cell motility in ST6Gal-I-KD HCT116 cells.