Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiles of the human breast cancer cell line MDA-MB-231 co-cultured with adipose tissue from mice on chow or high-fat diet

ABSTRACT: Obesity is a known risk factor for breast cancer. To identify genes and underlying pathways in human triple-negative breast cancer cells affected by interaction with adipose tissue, MDA-MB-231 breast cancer cells were cultivated in a co-culture system with or without adipose tissue explants from mice for the purpose of a microarray gene expression analysis. Co-culture of MDA-MB-231 breast cancer cells was performed with adipose tissue explants obtained from C57BL/6J mice that were fed a high-fat diet (HFD, 58%Kcal from fat) or normal chow diet (NC; 11%Kcal from fat) ad libitum for 16 weeks. For co-cultivation analyses of breast cancer cells and adipose tissue explants, we set up a two-dimensional transwell system, which enables intercellular communication through soluble factors secreted into the medium but inhibits intermixture of the different cell types. Following 72 hours of co-culture with or without adipose tissue, total RNA was isolated from the breast cancer cells and subjected to microarray gene expression analyses.

ORGANISM(S): Homo sapiens

SUBMITTER: Ralph Burkhardt

PROVIDER: E-MTAB-9485 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Inhibition of cannabinoid receptor 1 (CB1) has shown efficacy in reducing body weight and improving metabolic parameters, with the effects correlating with target engagement in the brain. Recently, the peripheral effects of inhibiting the CB1 receptor has been appreciated through studies in diet-induced obese and liver-specific CB1 KO mice. In this report, we systematically investigated gene expression changes in peripheral tissues of DIO mice treated with the CB1 inverse agonist AM251. CB1 receptor inhibition led to down-regulation of genes within the de novo fatty acid and cholesterol synthetic pathways, including SREBP-1 and -2, and their downstream targets in both liver and adipose tissue. In addition, genes involved in fatty acid Beta-oxidation were up-regulated with AM251 treatment, probably through the activation of PPARalpha. In adipose tissue, CB1 receptor inhibition led to the down-regulation of genes in the TNFalpha signal transduction pathway and possibly to the activation of PPARgamma, both of which would result in improved insulin sensitivity. CB1-/- mice were obtained from A. Zimmer (University of Bonn) (Zimmer et al., 1999) and back-crossed onto C57BL/6J genetic background for ten generations by A. Zimmer before homozygous CB1-/- mice were re-derived at Taconic Farms (German Town, NY) onto the C57BL/6N genetic background. Male CB1-/- mice and control littermates (n=5-7 in each group) at 2 months of age were fed with regular chow (Teklad 7012, 13 % kcal from fat, 3.41 kcal/g, Harlan Laboratories, Indianapolis, IN, USA) or high fat diet (S3282, 59.4% kcal from fat; 24.5% kcal from carbohydrate; 16.2% kcal from protein; 5.29 kcal/g, Bio-Serv, Frenchtown, NJ, USA) for 14 weeks, and were individually caged 1 week before drug treatment. Vehicle (0.5% methylcellulose) or AM251 (10 mg/kg, Sigma, St Louis, MO, USA) were dosed by oral gavage at 5:00 PM daily for 2 days. Body weight was measured at 5:00 PM on day 1 and 2, and at 10:00 AM on day 3 before tissue collection. Food was measured at 5:00 PM on day 1 and at 10:00 AM on day 3 before tissue collection. Food intake is calculated as the difference in food weight at the start minus at the end of the study. Mice were euthanized by CO2 asphyxiation at 10:00 AM following the second dose.

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Gene expression profiles of the human breast cancer cell line MDA-MB-231 co-cultured with adipose tissue from mice on chow or high-fat diet

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