Study on the anti-invasion effect of SEPT7 gene for U251MG glioma cell in vitro.
- Author:
Song XU
1
;
Zhi-fan JIA
;
Qiang HUANG
;
Chunsheng KANG
;
Guang-xiu WANG
;
An-ling ZHANG
;
Xiao-zhi LIU
;
Xuan ZHOU
;
Peng XU
;
Pei-yu PU
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Blotting, Western; Cell Cycle Proteins; genetics; physiology; Cell Line, Tumor; Cell Movement; genetics; Genetic Vectors; genetics; Glioma; metabolism; pathology; Humans; Integrin alphaVbeta3; metabolism; Matrix Metalloproteinase 14; metabolism; Matrix Metalloproteinase 2; metabolism; Matrix Metalloproteinase 9; metabolism; Microscopy, Confocal; Neoplasm Invasiveness; genetics; Septins; Tissue Inhibitor of Metalloproteinase-1; metabolism; Tissue Inhibitor of Metalloproteinase-2; metabolism
- From:Chinese Journal of Medical Genetics 2008;25(3):262-267
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the anti-invasion effect of SEPT7 gene on U251MG glioma cells and its possible molecular mechanism.
METHODSRecombinant adenovirus vector carrying SEPT7 gene (rAd5-SEPT7) was transduced to human glioma cell line U251MG, and empty adenovirus vector was used as control. Tumor invasion was examined by Transwell method and 3 D-Matrigel assay, and tumor cell migration by wound-healing method and 2 D-Matrigel assay. Three major molecular events associated with cell motility and migration, including changes of expression in MMP2, MMP9, MT1-MMP, TIMP1 and TIMP2, the alteration of integrin alpha(v)beta(3) expression, and the structural change of cytoskeleton protein, tubulin-alpha, in U251 cells transduced with rAd5-SEPT7 were studied by Western blotting, immunofluorescence and laser scanning confocal microscope, respectively.
RESULTSThe invasive and migratory capabilities of cells transduced with rAd5-SEPT7 were inhibited. The expression of extracellular matrix metalloproteinases MMP-2, MMP-9, MT1-MMP and integrin alpha(v)beta(3) was significantly decreased, while the expression of matrix metalloproteinase inhibitor TIMP1, TIMP2 was upregulated. Intracellular cytoskeleton protein-tubulin-alpha in U251 cells exhibited prominent morphological changes which including the appearance of distortion and aggregation resulting from redistribution of tubulin-alpha, and this feature of alteration was similar to the tubulin-alpha structure in normal non-tumor cells.
CONCLUSIONSEPT7 gene can inhibit the invasion and migration ability of U251 glioma cells. Its molecular mechanism may include that SEPT7 gene reverses the imbalanced state of MMPs/TIMPs, downregulates the expression of integrin alpha(v)beta(3) and alters the structure of tubulin-alpha of U251MG glioma cells. It is suggested that SEPT7 gene could be a good candidate for gene therapy of gliomas.
