Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: IGFBP2 regulated genes in BT474 breast cancer cells. a ) Western blot analysis of IGFBP2 in the supernatant of IGFBP2 knockdown clones C5 and C12 and control cells. Lower panel is ponceau stained membrane shown as loading control. b ) Hierarchical cluster of differentially expressed genes in IGFBP2 knockdown BT474 cells versus control cells. Differentially regulated genes were clustered using MeV software. The dendrogram on the left shows different clusters of genes segregated according to the pattern of regulation. Red and green indicate high and low expression of genes respectively. Black indicates no regulation. c ) Validation of selected genes by qPCR. Bar graphs of differentially regulated genes in IGFBP2 Knockdown BT474 clones versus control cells. The graphs represent the fold change over control after normalization with the expression of RPL35a.

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Western Blot, Knockdown, Clone Assay, Control, Staining, Membrane, Software, Expressing, Biomarker Discovery

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 differentially regulated genes (p < 0.05) in IGFBP2 positive versus IGFBP2 negative tumors

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques:

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: Differential expression of genes in IGFBP2 positive and IGFBP2 negative tumor samples compared to control tissues. a ) Genes were clustered using MeV software. The dendrogram on the left shows different clusters of genes segregated according to the pattern of regulation. Red and green indicate high and low expression of genes respectively. Black indicates no regulation. b ) Validation of selected genes. Scatter plots of differentially regulated genes in IGFBP2 positive and IGFBP2 negative tumors compared to the expression in normal tissues. Log 2-transformed gene expression ratios obtained from real-time quantitative PCR analysis normalized to TBP expression are plotted. Each dot represents a data derived from one sample.

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Quantitative Proteomics, Control, Software, Expressing, Biomarker Discovery, Transformation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 common genes differentially regulated between IGFBP2 positive tumors and IGFBP2 knockdown clones

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Knockdown

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: β-catenin and IGFBP2 expression in IGFBP2 knockdown and control cells. IGFBP2 knockdown clones and control cells were plated on coverslips and allowed to grow. 24 h after plating, cells were fixed, permeabilized, and stained for β-catenin and IGFBP2 expression. Expression of β-catenin and IGFBP2 is shown in green and blue, respectively. Nucleus was stained using propidium iodide (PI) as shown in red. Original magnification was 63×.

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Expressing, Knockdown, Control, Clone Assay, Staining

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: Regulation of β-catenin by IGFBP2 is IGF1R and FAK dependent. IGFBP2 knockdown clone C12 was transfected with IGFBP2 and 36 h. after transfection cells were fixed and analyzed for β-catenin, and IGFBP2. For inhibitor experiments, 24 h. after transfection cells were treated with IGF1R or FAK inhibitor for 12 h. Cells were fixed and analyzed for β-catenin and IGFBP2 expression. Expression of β-catenin and IGFBP2 is shown in green and blue, respectively. Nucleus was stained using propidium iodide (PI) as shown in red. Original magnification was 63×. V, Vector control; OE, Over expression; Inh, inhibitor.

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Knockdown, Transfection, Expressing, Staining, Plasmid Preparation, Control, Over Expression

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: β-catenin and IGFBP2 expression in breast cancer tissues. Representative micrographs showing β-catenin staining in cancer tissues. a β-catenin staining of a section of IDC 3 breast tumors showing cytoplasmic and membrane staining. b represents breast cancer tissue section stained for IGFBP2 showing predominantly cytoplasmic staining.

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Expressing, Staining, Membrane

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: IGFBP2 and β-catenin expression in breast cancer tissues

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Expressing

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 differentially regulated genes (p < 0.05) in IGFBP2 knockdown clones

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques: Knockdown, shRNA

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: GSEA summary of pathways associated with genes down regulated upon IGFBP2 knockdown

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques:

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: GSEA summary of pathways associated with genes up regulated in IGFBP2 positive tumors

Article Snippet: In brief, breast cancer cells were transfected with IGFBP2 shRNA expression vector (Origene, Cat no. TR316590) or empty vector (Origene, Cat no. TR20003) and 48 hrs after transfection puromycin (1 μg/ml, Calbiochem) was added to the growth medium.

Techniques:

Journal: Oncotarget

Article Title: IGFBP2 plays an important role in heat shock protein 27-mediated cancer progression and metastasis

doi: 10.18632/oncotarget.18989

Figure Lengend Snippet: The silencing of Hsp27 decreased the IGFBP2 ( A ) mRNA and protein levels in HepJ5 cells. ( B ) The expression correlation of Hsp27 and IGFBP2 was found in tissue array. The overexpression of IGFBP2 partially reversed the deficits in proliferation ( C ), migration and invasion ( D ) in Hsp27-KD HepJ5 cells.

Article Snippet: The Human IGFBP2 overexpression plasmid (RC202573) was purchased from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: Expressing, Over Expression, Migration

Journal: Oncotarget

Article Title: IGFBP2 plays an important role in heat shock protein 27-mediated cancer progression and metastasis

doi: 10.18632/oncotarget.18989

Figure Lengend Snippet: ( A ) IGGBP2 protein expression in five HCC cell lines. (A) A ChIP assay of the IGFBP2 promoter region was performed using the Hsp27 antibody in HepJ5 and HepJ5 Hsp27-KD cells. Resveratrol, which induces PPARγ activation at the IGFBP2 promoter region, was used as a positive control. The IGFBP2 promoter region from −732 to −343 was amplified by PCR from the eluents from each group and analyzed by agarose gel electrophoresis. ( B ) Real-time PCR (Q-PCR) quantitative curve of the eluents from the IGFBP2 ChIP assay. IGFBP2 promoter fragments were measured and compared by Q-PCR analysis. ( C ) The IGFBP2 luciferase assay used the PGL3 vector containing the IGFBP2 promoter region from −1265 to +465. Post IGFBP2-PGL3 transfection on Hep-J5 and Hep-J5 Hsp27KD cells, the luciferase activities were measured and normalized with RLTK activity. The luciferase activity from empty vector was defined as a one-fold. *indicates p < 0.05. ( D ) Schematic of Hsp27 regulation in HCC.

Article Snippet: The Human IGFBP2 overexpression plasmid (RC202573) was purchased from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: Expressing, Activation Assay, Positive Control, Amplification, Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction, Luciferase, Plasmid Preparation, Transfection, Activity Assay