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

Authors Jiang L, Liang QL, Liang WM, Zhang HJ, Huang J, Yuan GL, Peng XX, Cheng SA, Huang ZG, Zhang XN

Received 5 June 2018

Accepted for publication 21 August 2018

Published 9 October 2018 Volume 2018:11 Pages 6665—6671

DOI https://doi.org/10.2147/OTT.S176388

Checked for plagiarism Yes

Review by Single-blind

Peer reviewers approved by Dr Colin Mak

Peer reviewer comments 2

Editor who approved publication: Dr Carlos E Vigil


Liang Jiang,1,* Qi-Lian Liang,2 Wei-Ming Liang,1,* Hui-Jie Zhang,2 Jie Huang,2 Gao-Le Yuan,2 Xiao-Xia Peng,2 Shao-Ang Cheng,2 Zhi-Gang Huang,3 Xiang-Ning Zhang4

1Interventional Ward, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China; 2Oncology Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China; 3Department of Epidemiology, School of Public Health, Guangdong Medical University, Dongguan 523808, China; 4Department of Pathophysiology, School of Basic Medical Science, Guangdong Medical University, Dongguan 523808, China

*These authors contributed equally to this work

Introduction: 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.
Materials and methods: 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.
Result: 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 P<0.001). DNM3 downregulated the protein expression of MMP-2 and MMP-9.
Conclusion: DNM3 may weaken the malignant behavior of colon cancer and may have promoted the invasion and migration of colon cancer by regulating the expression of MMP-2 and MMP-9.

Keywords: dynamin 3, colon cancer, proliferation, migration, invasion

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