School of Life Science and Medicine, Dalian University of Technology, Panjin, China.
Department of Biochemistry, Dalian Medical University, Dalian, China.
Sci Rep. 2018 May 9;8(1):7366. doi: 10.1038/s41598-018-25768-z.
MicroRNAs (miRNAs) and aberrant glycosylation both play important roles in tumor metastasis. In this study, the role of miR-23a in N-glycosylation and the metastasis of mouse hepatocellular carcinoma (HCC) cells was investigated. The miRNA expression array profiles that were confirmed by qPCR and Western blot analyses revealed higher miR-23a expression levels in Hca-P cells (with lymphatic metastasis potential) than in Hepa1-6 cells (with no lymphatic metastasis potential), while the expression of mannoside acetylglucosaminyltransferase 3 (Mgat3) was negatively associated with metastasis potential. Mgat3 is a key glycosyltransferase in the synthesis of the bisecting (β1,4GlcNAc branching) N-glycan structure. Bioinformatics analysis indicated that Mgat3 may be a target of miR-23a, and this hypothesis was verified by dual-luciferase reporter gene assays. Furthermore, we found that the transcription factor Runx2 can directly bind to the miR-23a gene promoter and promote its expression, as shown in dual-luciferase reporter gene assays and ChIP assays. Collectively, these results indicate that miR-23a might increase the metastatic potential of mouse HCC by affecting the branch formation of N-glycan chains presented on the cell surface through the targeting of the glycosyltransferase Mgat3. These findings may provide insight into the relationship between abnormal miRNA expression and aberrant glycosylation during tumor lymphatic metastasis.
微小 RNA(miRNA)和异常糖基化在肿瘤转移中都发挥着重要作用。本研究探讨了 miR-23a 在 N-糖基化和小鼠肝癌(HCC)细胞转移中的作用。qPCR 和 Western blot 分析证实的 miRNA 表达谱显示,具有淋巴转移潜能的 Hca-P 细胞中 miR-23a 的表达水平高于无淋巴转移潜能的 Hepa1-6 细胞,而甘露糖苷乙酰氨基葡萄糖基转移酶 3(Mgat3)的表达与转移潜能呈负相关。Mgat3 是合成双分支(β1,4GlcNAc 分支)N-糖链结构的关键糖基转移酶。生物信息学分析表明,Mgat3 可能是 miR-23a 的靶基因,双荧光素酶报告基因实验验证了这一假设。此外,我们发现转录因子 Runx2 可以直接结合 miR-23a 基因启动子并促进其表达,这在双荧光素酶报告基因实验和 ChIP 实验中得到了证实。综上所述,这些结果表明,miR-23a 可能通过靶向糖基转移酶 Mgat3 影响细胞表面 N-糖链分支形成,从而增加小鼠 HCC 的转移潜能。这些发现可能为异常 miRNA 表达与肿瘤淋巴转移过程中异常糖基化之间的关系提供新的认识。