南方医科大学学报2018，Vol.38Issue(1)：14-19,6.
过表达DLL3促进人胃癌细胞的增殖
Over-expression of human Notch ligand Delta-like 3 promotes proliferation of human gastric cancer cells in vitro
摘要
Abstract
Objective To construct a eukaryotic expression plasmid carrying human full-length Notch ligand Delta-like 3 (DLL3) gene and study the effect of DLL3 knockdown and overexpression on the proliferation of gastric cancer cells in vitro. Methods Human full-length DLL3 gene was amplified by PCR and cloned into the eukaryotic expression vector pCMV-Tag4. After verification by restriction enzymes and sequencing, the recombinant DLL3/pCMV-Tag4 vector was transiently transfected into HEK293T cells, in which the expressions of human DLL3 mRNA and protein were detected using real-time quantitative PCR and Western blotting, respectively. The expression of DLL3 in normal gastric epithelial cells and gastric cancer cell lines was detected by qRT-PCR and Western blotting. DLL3/pCMV-Tag4 was transfected into 3 gastric cancer cell lines, and their proliferation was assessed with MTT assay. Human gastric cancer cells MGC803 and MKN45 were also transfected with a specific human DLL3-siRNA to assess the effect of DLL3 down-expression on the cell proliferation. Results The recombinant eukaryotic expression vector DLL3/pCMV-Tag4 was successfully constructed and human full-length DLL3 was expressed in HEK293T cells. MTT assay showed that DLL3 over-expression obviously promoted the proliferation and down-regulation of DLL3 inhibited the proliferation of the gastric cancer cells. Conclusion DLL3 overexpression can promote the proliferation of gastric cancer cells in vitro, and down-regulation of DLL3 inhibits the proliferation of gastrc cancer cells,which provides a novel strategy for targeted thrapy of gastric cancer.关键词
人DLL3/真核表达/胃癌细胞/增殖Key words
human Delta-like ligand 3/eukaryotic expression/gastric cancer cell line/proliferation引用本文复制引用
胡冰心,宁云山,叶健斌,邱晓媚,林彦青,吴丹琳,温俊杰,罗美群,宁立军,李妍..过表达DLL3促进人胃癌细胞的增殖[J].南方医科大学学报,2018,38(1):14-19,6.基金项目
国家自然科学基金(81470831) （81470831）
国家自然科学基金-联合项目(U1401223) （U1401223）
广东省科技计划项目(2013B090800036,2015A040404021, 2016B090919019) （2013B090800036,2015A040404021, 2016B090919019）
广东省大学生创新创业项目(201512121122) （201512121122）
广东省大学生创新训练项目(201612121100,201612121243,201612121034) Supported by National Natural Science Foundation of China (81470831, U1401223). （201612121100,201612121243,201612121034）
