Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Cell proliferation of human breast cancer cell lines . (a) MDAMB231 and (b) MDAMB468. Cells were treated with increasing concentrations of lovastatin lactone or lovastatin acid (μg/mL) for 48 hours. Data are presented as mean ± standard deviation (n = 5) * P < 0.05;** P < 0.05; *** P < 0.001.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Standard Deviation

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Changes in expression of proteins involved in (a) regulation of cell cycle and cell death and (b) oxidative and metabolic processes of human MDAMB231 and MDAMB468 cells . Both cell lines were treated with 8 μg/mL lovastatin lactone (Lova Lac) or lovastatin acid (Lova Ac) for 48 hours. Data represent relative spot volumes (as calculated from two-dimensional gel images of whole cell extracts; data are presented as mean ± standard deviation (n = 5) * P < 0.05;** P < 0.05; *** P < 0.001). Gel spots which showed significant differences in their volume between the control and lovastatin-treated cells were cut-out, proteins were digested and analyzed using liquid chromatography (LC) mass spectrometry (MS)/MS analysis. In MDAMB231 cells they were identified as GTPase-activating protein SH3-domain-binding protein 1 (G3BP1), TNF type 1 receptor-associated protein (TRAP1) and glutathione S-transferase (GST) omega proteins (Table 1a), whereas the spot belonging to citrate lyase beta and sterol carrier protein 2 (SCP-2) originated from MDAMB468 cells (Table 1b). Cofilin1/2 was identified as upregulated in both cell lines. The image and changes as observed in MDAMB231 cells is shown.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Expressing, Two-Dimensional Gel Electrophoresis, Standard Deviation, Control, Liquid Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Binding Assay

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Western blot analysis of proteins involved in small GTPase-mediated cell signaling . Breast cancer cell lines MDAMB231 and MDAMB468 were treated with 8 μg/mL lovastatin lactone (Lova Lac) or lovastatin acid (Lova Ac) for 48 hours. For key proteins, western blot analysis was performed based on MDAMB231 cell extracts (for Ras homolog gene family member A (RhoA), cell division cycle 42 (CDC42) and GTPase-activating protein SH3-domain-binding protein 1 - phospho form (pG3BP1)), otherwise both cell lines are shown. Densitometry data were normalized based on the amount of β-actin. Data are presented as means ± standard deviations (n = 3) * P < 0.05;** P < 0.05; *** P < 0.001). Gel images were cropped to improve the clarity and conciseness of the presentation. GDI-2, Rho GDP dissociation inhibitor 2.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Western Blot, Binding Assay

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Western blot analysis of proteins involved in regulation of the cell cycle including the modulation of the E2F1-Rb activity . Breast cancer cell lines MDAMB231 and MDAMB468 were treated with 8 μg/mL lovastatin lactone (Lova Lac) or lovastatin acid (Lova Ac) for 48 hours. Western blot analysis of prohibitin was performed based on MDAMB231 cell extracts, otherwise both cell lines are shown. Densitometry data were normalized based on the amount of β-actin. Data are presented as means ± standard deviations (n = 3) * P < 0.05;** P < 0.05; *** P < 0.001. Gel images were cropped to improve the clarity and conciseness of the presentation. HMGB1, high-mobility group box 1; MCM7, minichromosome maintenance protein 7; MSH2, MutS homolog 2.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Western Blot, Activity Assay

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Western blot analysis of proteins involved in regulation of apoptosis and AKT-signaling . Breast cancer cell lines MDAMB231 and MDAMB468 were treated with 8 μg/mL lovastatin lactone (Lova Lac) or lovastatin acid (Lova Ac) for 48 hours. For key proteins, western blot analysis of phosphatase and tensin homolog (PTEN), pAkt and N-myc downstream regulated gene 1 (NDRG1) was performed based on MDAMB231 cell extracts, otherwise both cell lines are shown. Densitometry data were normalized based on the amount of β-actin. Data are presented as means ± standard deviations (n = 3) * P < 0.05;** P < 0.05; *** P < 0.001). Gel images were cropped to improve the clarity and conciseness of the presentation. PCNA, proliferating cell nuclear antigen.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Intracellular concentrations (nmol/g cell weight) of 13 C-labeled endogenous metabolites (glycolysis and TCA cycle intermediates, glucose) and lipid metabolites (choline-containing phospholipids, cholesterol)

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Changes in intracellular 13 C-labeled -alanine, -lactate, -glucose and -glutamine signals in MDAMB468 cells treated with 8 μg/mL lovastatin acid for 48 hours . 13 C-NMR spectra with embedded, corresponding 1 H-NMR spectra are shown (including citrate at 2.52 + 2.69 ppm). Arrows indicate the direction of signal changes (increase or decrease). Ala, alanine; Gln, glutamine; Glu, glutamine; GSH, total glutathione; Lac, lactate.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Labeling

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Representative 1 H-NMR spectra of MDAMB468 lipid extracts . Cells were treated with 8 μg/mL lovastatin acid for 48 hours. Arrows indicate the direction of signal changes (decrease). Chol, cholesterol (C18 and C19, CH 3 ); Δ (δ), double bond; F, fatty acid side chain; F α , F β , protons in the fatty acid chain; Fmix: -(CH 2 )n-, tCho, total choline-containing phospholipids.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: Effects of lovastatin on breast cancer cells: a proteo-metabonomic study

doi: 10.1186/bcr2485

Figure Lengend Snippet: Schematic diagram summarizing the effects of lovastatin lactone and acid on signaling pathways as found in the present study . The solid arrows mark the directional change of proteins (up- or down-regulation). Doted arrows mark hypothesized change in protein expression/activity. CDC42, cell division cycle 42; G3BP1, GTPase-activating protein SH3-domain-binding protein 1; GDI-2, Rho GDP dissociation inhibitor 2; LIMK, LIM domain kinase; MAPK, mitogen-activated protein kinase; MCM7, minichromosome maintenance protein 7; MSH2, MutS homolog 2; NDRG1, N-myc downstream regulated gene 1; p21, cyclin-dependent kinase inhibitor 1A; PCNA, proliferating cell nuclear antigen; PI3K, phosphoinositide 3-kinase; PTEN, phosphatase and tensin homolog; Rb, retinoblastoma protein; RhoA, Ras homolog gene family member A.

Article Snippet: Lovastatin (in its lactone and hydroxy acid form) was purchased from Toronto Research Chemicals (North York, Ontario, Canada).

Techniques: Protein-Protein interactions, Expressing, Activity Assay, Binding Assay