Furong Li1, Xinghua Zhou2, Ming Chen2, Wei Fan3. 1. Department of Laboratory medicine, Shanghai University of Medicine & Health Sciences Affiliated Zhoupu Hospital, Shanghai 201318, China; Department of Immunology, Zunyi Medical University, Zunyi 563099, Guizhou, , China. 2. Department of Laboratory medicine, Shanghai University of Medicine & Health Sciences Affiliated Zhoupu Hospital, Shanghai 201318, China. 3. Department of laboratory medicine, Huaian Tumor Hospital, No.14 Yue miao East Street, Huaian 223200, Jiangsu Province, China. Electronic address: wei202005fan@163.com.
Abstract
OBJECTIVE: This experiment will explore the effect of LncRNA DRAIC/miR-149-5p/NFIB molecular network on esophageal cancer (EC) cells' biological behavior and autophagy. METHODS: We bought human EC cells and normal esophageal epithelial cells HEEC. DRAIC, miR-149-5p and NFIB protein expression were tested. The low expression plasmid of DRAIC and empty vector of DRAIC, miR-149-5p miR-mimics, miR-149-5p inhibitors and negative control groups, NFIB high expression plasmid, NFIB low expression plasmid and empty vector were transfected into EC cells (Eca-109 and EC9706) to detect changes in cell biological behavior and autophagy protein expression. The targeted relationship between DRAIC/miR-149-5p/NFIB was verified through dual-luciferase report and pull-down experiment. RESULTS: DRAIC and NFIB showed high expression in EC cells, while miR-149-5p showed low expression. Down-regulating DRAIC, NFIB and over-expressing miR-149-5p can inhibit EC cells' proliferation and invasion, and improve apoptosis and autophagy. Dual-luciferase report and pull-down experiment confirmed that DRAIC targeted miR-149-5p regulation, and down-regulating DRAIC could reverse miR-149-5p inhibitor's effect on the biological behavior of EC cells. However, dual-luciferase report revealed that miR-149-5p directly targeted NFIB, and miR-149-5p inhibitor could weaken the effect of down-regulating NFIB on apoptosis and autophagy of EC cells. Moreover, DRAIC has an effect on the autophagy of EC cells through miR-149-5p/NFIB. CONCLUSION: LncRNA DRAIC is relevant to cell biology and autophagy of EC. In the future, DRAIC may be developed as a key gene for EC diagnosis and treatment.
OBJECTIVE: This experiment will explore the effect of LncRNA DRAIC/miR-149-5p/NFIB molecular network on esophageal cancer (EC) cells' biological behavior and autophagy. METHODS: We bought human EC cells and normal esophageal epithelial cells HEEC. DRAIC, miR-149-5p and NFIB protein expression were tested. The low expression plasmid of DRAIC and empty vector of DRAIC, miR-149-5p miR-mimics, miR-149-5p inhibitors and negative control groups, NFIB high expression plasmid, NFIB low expression plasmid and empty vector were transfected into EC cells (Eca-109 and EC9706) to detect changes in cell biological behavior and autophagy protein expression. The targeted relationship between DRAIC/miR-149-5p/NFIB was verified through dual-luciferase report and pull-down experiment. RESULTS:DRAIC and NFIB showed high expression in EC cells, while miR-149-5p showed low expression. Down-regulating DRAIC, NFIB and over-expressing miR-149-5p can inhibit EC cells' proliferation and invasion, and improve apoptosis and autophagy. Dual-luciferase report and pull-down experiment confirmed that DRAIC targeted miR-149-5p regulation, and down-regulating DRAIC could reverse miR-149-5p inhibitor's effect on the biological behavior of EC cells. However, dual-luciferase report revealed that miR-149-5p directly targeted NFIB, and miR-149-5p inhibitor could weaken the effect of down-regulating NFIB on apoptosis and autophagy of EC cells. Moreover, DRAIC has an effect on the autophagy of EC cells through miR-149-5p/NFIB. CONCLUSION: LncRNA DRAIC is relevant to cell biology and autophagy of EC. In the future, DRAIC may be developed as a key gene for EC diagnosis and treatment.