Suppression of cellular O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) can repress proliferation and migration of various cancer cells, which opens a new avenue for cancer therapy. Based on the regulation of insulin gene transcription, we designed a cell-based fluorescent reporter capable of sensing cellular O-GlcNAcylation in HEK293T cells. The fluorescent reporter mainly consists of a reporter (green fluorescent protein (GFP)), an internal reference (red fluorescent protein), and an operator (neuronal differentiation 1), which serves as a “sweet switch” to control GFP expression in response to cellular O-GlcNAcylation changes. The fluorescent reporter can efficiently sense reduced levels of cellular O-GlcNAcylation in several cell lines. Using the fluorescent reporter, we screened 120 natural products and obtained one compound, sesamin, which could markedly inhibit protein O-GlcNAcylation in HeLa and human colorectal carcinoma-116 cells and repress their migration in vitro. Altogether, the present study demonstrated the development of a novel strategy for anti-tumor drug screening, as well as for conducting gene transcription studies.

抑制细胞 O-连接的 β- N-乙酰氨基葡糖化（O-GlcNAcylation）可以抑制各种癌细胞的增殖和迁移，为癌症治疗开辟了新途径。基于胰岛素基因转录的调控，我们设计了一种基于细胞的荧光报告基因，能够感知 HEK293T 细胞中的细胞 O-GlcNAcylation。荧光报告基因主要由报告基因（绿色荧光蛋白（GFP））、内参（红色荧光蛋白）和操作符（神经元分化 1）组成，作为“甜蜜开关”控制 GFP 表达以响应细胞 O-GlcNAcylation 改变。荧光报告基因可以有效地检测几种细胞系中细胞 O-GlcNAcylation 水平的降低。使用荧光报告基因，我们筛选了 120 种天然产物，得到了一种化合物芝麻素，可显着抑制HeLa和人结直肠癌116细胞中的蛋白质O-GlcNAcylation并抑制其体外迁移。总之，本研究证明了一种用于抗肿瘤药物筛选以及进行基因转录研究的新策略的发展。