A living cell-based fluorescent reporter for high-throughput screening of anti-tumor drugs
- Author:
Ningning TANG
1
;
Ling LI
;
Fei XIE
;
Ying LU
;
Zifan ZUO
;
Hao SHAN
;
Quan ZHANG
;
Lianwen ZHANG
Author Information
1. College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research,Nankai University,Tianjin,300350,China
- Keywords:
Fluorescent reporter;
High-throughput screening;
O-linked β-N-acetylglucosaminylation;
Anti-tumor drug;
Gene transcriptional regulation
- From:
Journal of Pharmaceutical Analysis
2021;11(6):808-814
- CountryChina
- Language:Chinese
-
Abstract:
Suppression of cellular O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) can repress prolifera-tion 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 con-ducting gene transcription studies.
