Cytoskeleton-associated Genes Research Articles

Global Gene Expression Analysis Identifies PDEF Transcriptional Networks Regulating Cell Migration during Cancer Progression

Prostate derived ETS factor (PDEF) is an ETS (epithelial-specific E26 transforming sequence) family member that has been identified as a potential tumor suppressor. In multiple invasive breast cancer cells, PDEF expression inhibits cell migration by preventing the acquisition of directional morphological polarity conferred by changes in cytoskeleton organization. In this study, microarray analysis was used to identify >200 human genes that displayed a common differential expression pattern in three invasive breast cancer cell lines after expression of exogenous PDEF protein. Gene ontology associations and data mining analysis identified focal adhesion, adherens junctions, cell adhesion, and actin cytoskeleton regulation as cell migration-associated interaction pathways significantly impacted by PDEF expression. Validation experiments confirmed the differential expression of four cytoskeleton-associated genes with known functional associations with these pathways: uPA, uPAR, LASP1, and VASP. Significantly, chromatin immunoprecipitation studies identified PDEF as a direct negative regulator of the metastasis-associated gene uPA and phenotypic rescue experiments demonstrate that exogenous urokinase plasminogen activator (uPA) expression can restore the migratory ability of invasive breast cancer cells expressing PDEF. Furthermore, immunofluorescence studies identify the subcellular relocalization of urokinase plasminogen activator receptor (uPAR), LIM and SH3 protein (LASP1), and vasodilator-stimulated protein (VASP) as a possible mechanism accounting for the loss of morphological polarity observed upon PDEF expression.

Gene expression profiles in the liver of mice irradiated with 60Co gamma rays and treated with soybean isoflavone

To better understand the molecular mechanisms underlying the radio-protective effect of soybean isoflavone that we observed in our recent animal experiments. We utilized a cDNA microarray to investigate the expression profiles of 4,096 known genes in the livers of irradiated-mice with or without soybean isoflavone treatment. Dye swap approach was employed to control for gene-specific dye bias and quantitative real-time RT-PCR was performed on several genes to validate the cDNA microarray data. Compared with the control group, 68 genes were up-regulated and 28 genes were down-regulated in mice treated with irradiation alone, whereas only 6 genes were down-regulated and 35 genes were up-regulated in mice treated with soybean isoflavone. Interestingly, some of the down-regulated genes in the irradiated group, such as DNA repair and stress response genes and cytoskeleton-associated genes, which are markers of cellular damage after irradiation, were maintained at close to normal expression levels after soybean isoflavone treatment. Comparison of gene expression profiles in the livers of irradiated-mice treated with or without soybean isoflavone suggested that soybean isoflavone may be an efficient tool to reverse irradiation damage of the liver through multiple-pathways and also provides important clues to further pursue the molecular mechanisms underlying the radio-protective activity of soybean isoflavone.

Effect of Helicobacter pylori VacA on gene expression of gastric cancer cells.

To determine the effect of Helicobacter pylori VacA on gene expression of gastric cancer cells. Gene expression profile of a gastric cancer cell line, SGC7901, after challenged by VacA+ and VacA- H pylori broth culture supernatants (BCS), was detected by the cDNA microarray technique. Cytoskeleton changes of SGC7901 and HeLa cells were observed through high-resolution laser scanning confocal microscopy. A total of 16,000 cDNA clones were detected. The percentage of genes with heterogeneous expression in SGC7901 cells challenged by VacA+ BCS reached 5%, compared with that challenged by VacA- BCS. There were 865 genes/EST with 2-fold differential expression levels and 198 genes/EST with 3-fold differential expression levels. Most of these genes were involved in vital cell events including signal transduction, regulation of gene expression, cytoskeleton, apoptosis, stress response and inflammation, cell cycle and tumor development. Cells co-cultured with VacA+ BCS showed collapsed and disrupted microtubular cytoarchitecture. VacA+ BCS can disrupt cytoskeletal architecture, likely through affecting the expression of cytoskeleton-associated genes, directly induce the expression of tumor promoter-related genes and inhibit the expression of tumor suppressor genes, thus favoring the development of tumors. VacA+ BCS can also alter the expression of inflammation and stress response genes. This suggests that VacA may play an important role in the pathogenicity of H pylori.

Analysis of gene expression profile in gastric cancer cells stimulated with Helicobacter pylori isogenic strains.

To understand the biological processes within host cells induced by VacA, isogenic strains of Helicobacter pylori (NCTC 11638 or 11638-DeltavacA) were used to stimulate gastric cancer cells SGC7901, and differentially expressed genes in host cells were identified using cDNA microarray technology. More than 300 genes were found to alter their mRNA expression at different time points, among which 68 were related to the cytoskeleton, 87 were associated with cell cycle, cell death and proliferation, IL8 expression was also found to be up-regulated. Cells co-cultured with broth-culture supernatant (BCS) of NCTC 11638 showed more alteration in microtubule cytoskeleton morphology, as observed by laser scanning confocal microscopy, and a lower apoptosis rate, detected by flow cytometry, compared with those co-cultured with BCS of 11638-DeltavacA. The supernatants of cells co-cultured with NCTC 11638 showed significantly higher IL8 expression than those co-cultured with 11638-DeltavacA. It is concluded that VacA disrupts cytoskeletal architecture by influencing the expression of cytoskeleton-associated genes. VacA breaks the balance between cell proliferation and cell death by inducing the maladjustment of genes related to cell cycle. VacA is also able to induce the inflammatory response.