过表达外源FUT8增加乳腺癌细胞的迁移和侵袭能力

Exogenetic Overexpression of FUT8 Increases the Ability of Migration and Invasion in Breast Cancer Cells

核心岩藻糖的修饰被认为是对糖蛋白进行转录后加工和功能调控的一种重要方式,与肿瘤的发生发展密切相关,但其在乳腺癌恶性转化过程中所发挥的生物学功能尚不明确.本文构建了α1,6-岩藻糖基移转酶(FUT8)基因真核表达载体,将其转染人乳腺癌细胞Bcap-37,实时PCR及Western印迹检测FUT8 mRNA和蛋白质表达,通过细胞划痕实验和Transwell小室观察FUT8过表达对细胞体外迁移、侵袭能力的影响,免疫共沉淀和Western印迹检测肿瘤细胞整合素、基质金属蛋白酶(matrix metalloproteinases,MMPs)家族相关蛋白质表达水平变化.结果显示,外源FUT8基因在mRNA水平和蛋白质水平的表达均显著增加,FUT8过表达组细胞迁移和侵袭能力明显增强;上调FUT8基因表达可使Bcap-37细胞中核心岩藻糖基化的整合素-α3β1、MMP-2和MMP-9在蛋白质水平呈不同程度升高.上述研究表明,FUT8可通过核心岩藻糖化修饰正性调控整合素-α3β1的功能,诱导MMP-2、MMP-9的表达,促进乳腺癌细胞的迁移和侵袭.

Modification of core fucosylation is regarded as an important manner of post-translational processing and functional regulation of glycoproteins, it was closely related to cancer tumorigenesis and progression. However, its specific biological functions in the process of breast cancer development remain unclear. In the current study, a FUT8 expression vector of eukaryote was constructed and transfected them into the human breast cancer Bcap-37 cells. After transient transfection, the expression of exogenous FUT8 was confirmed at the mRNA and protein levels by real-time PCR and Western blot assays, respectively. Cell scratch assay and transwell assays were used to determine cell migration and invasion. Integrins and matrix metalloproteinases （MMPs） protein levels were measured by immunoprecipitated and Western blotting. The results suggested that accompanied with the overexpression of FUT8 in transfected Bcap-37 cells, the abilities of migration and invasion were significantly increased in FUT8 overexpression cells, as compared to the original counterpart. FUT8 could up-regulates the protein expression of core fucosylated integrin-α3β1, MMP-2 and MMP-9 in Bcap-37 cells. This study showed that FUT8 can improve the invasion and migration ability of breast cancer cells through positive regulation core fucosylation of integrin-α3β1 and induce the expression of MMP-2 and MMP-9.