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.
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Subject:
Medicine, Pharmacy
>>
Preclinical Medicine
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Cite as:
ChinaXiv:201806.00161
(or this version
ChinaXiv:201806.00161V1)
DOI:10.12074/201806.00161V1
CSTR:32003.36.ChinaXiv.201806.00161.V1
- Recommended references:
胡冰心,叶健斌,邱晓媚,林彦青,吴丹琳,温俊杰,罗美群,宁立军,李妍,宁云山.(2018).过表达DLL3促进人胃癌细胞的增殖.南方医科大学学报.[ChinaXiv:201806.00161]
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