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

Abstract:

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.

journal_name

Onco Targets Ther

journal_title

OncoTargets and therapy

authors

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

doi

10.2147/OTT.S176388

subject

Has Abstract

pub_date

2018-10-09 00:00:00

pages

6665-6671

issn

1178-6930

pii

ott-11-6665

journal_volume

11

pub_type

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