Construction of a recombinant eukaryotic expression vector containing DNM3 gene and its expression in colon cancer cells

Dynamin 3 (DNM3) is a large GTPase that possesses mechanochemical properties and has been shown to be involved in malignancies. However, most studies about DNM3 are observational, and knowledge of the precise molecular mechanism of DNM3 remains limited. We constructed a PCDH-CMV-MCS-EF1a-GFP-Puro-DN...

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Bibliographic Details
Published in:OncoTargets and therapy 2018-01, Vol.11, p.6665-6671
Main Authors: Jiang, Liang, Liang, Qi-Lian, Liang, Wei-Ming, Zhang, Hui-Jie, Huang, Jie, Yuan, Gao-Le, Peng, Xiao-Xia, Cheng, Shao-Ang, Huang, Zhi-Gang, Zhang, Xiang-Ning
Format: Article
Language:English
Subjects:
Analysis
Anopheles
Apoptosis
Cancer cells
Cancer genetics
Cell adhesion & migration
Cloning
Colon cancer
Colorectal cancer
Deoxyribonucleic acid
DNA
DNA methylation
E coli
Genes
Genetic aspects
Genetic engineering
Genetic vectors
Liver cancer
Messenger RNA
Metastasis
Original Research
Plasmids
Proteins
RNA
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Summary:Dynamin 3 (DNM3) is a large GTPase that possesses mechanochemical properties and has been shown to be involved in malignancies. However, most studies about DNM3 are observational, and knowledge of the precise molecular mechanism of DNM3 remains limited. We constructed a PCDH-CMV-MCS-EF1a-GFP-Puro-DNM3 recombinant eukaryotic expression vector, which was then transfected into SW620 and LoVo cells. One cell line was divided into three groups. DNM3 mRNA and protein expression was analyzed by quantitative real-time PCR and Western blot assay. To investigate DNM3 biological activity in colon cancer SW620 and LoVo cell line, we performed cell proliferation, transwell migration, and invasion assay. Matrix metalloproteinase (MMP)-2 and MMP-9 protein expressions were detected by Western blot. We successfully constructed a PCDH-CMV-MCS-EF1a-GFP-Puro-DNM3 recombinant eukaryotic expression vector, and stable DNM3 expression was observed in SW620 and LoVo cell lines. The vector overexpressing DNM3 inhibited the proliferation, weak invasion, and migration ability of colon cancer SW620 and LoVo cells relative to those in the control group (all
ISSN:1178-6930
1178-6930
DOI:10.2147/OTT.S176388