Over-expression of human Notch ligand Delta-like 3 promotes proliferation of human gastric cancer cells in vitro
10.3969/j.issn.1673-4254.2018.01.003
- VernacularTitle:过表达DLL3促进人胃癌细胞的增殖
- Author:
Bingxin HU
1
;
Jianbin YE
;
Xiaomei QIU
;
Yanqing LIN
;
Danlin WU
;
Junjie WEN
;
Meiqun LUO
;
Lijun NING
;
Yan LI
;
Yunshan NING
Author Information
1. 南方医科大学检验与生物技术学院生物治疗研究所
- Keywords:
human Delta-like ligand 3;
eukaryotic expression;
gastric cancer cell line;
proliferation
- From:
Journal of Southern Medical University
2018;38(1):14-19
- CountryChina
- Language:Chinese
-
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.
