Effect of fatty acids from CLA-enriched egg yolk on MCF-7 adenocarcinoma cells
ABSTRACT: It was analyzed whole genome microarray data to describe the changes in gene transcription profile in human MCF-7 cancer cells under the influence of fatty acid extracts from CLA-enriched and non-enriched egg yolks.Those results might be found useful in assessing the application of CLA-enriched egg as a nutraceutics in cancer prevention. Six-condition experiment: CLA, KT, cis9,trans11-CLA, trans10,cis12-CLA, KT+cis9,trans11-CLA and KT+trans10,cis12-CLA vs MCF-7 adenocarcinoma cell line. Two control of expreriment. Three biological replicates and three technical replicates.
Project description:It was analyzed whole genome microarray data to describe the changes in gene transcription profile in human MCF-7 cancer cells under the influence of fatty acid extracts from CLA-enriched and non-enriched egg yolks.Those results might be found useful in assessing the application of CLA-enriched egg as a nutraceutics in cancer prevention. Overall design: Six-condition experiment: CLA, KT, cis9,trans11-CLA, trans10,cis12-CLA, KT+cis9,trans11-CLA and KT+trans10,cis12-CLA vs MCF-7 adenocarcinoma cell line. Two control of expreriment. Three biological replicates and three technical replicates.
Project description:In our previous study, we showed that fatty acids from CLA-enriched egg yolks (EFA-CLA) reduced the proliferation of breast cancer cells; however, the molecular mechanisms of their action remain unknown. In the current study, we used MCF-7 breast cancer cell line to determine the effect of EFA-CLA, as potential ligands for peroxisome proliferator-activated receptors (PPARs), on identified in silico PPAR-responsive genes: BCAR3, TCF20, WT1, ZNF621, and THRB (transcript TR?2). Our results showed that EFA-CLA act as PPAR ligands with agonistic activity for all PPAR isoforms, with the highest specificity towards PPAR?. In conclusion, we propose that EFA-CLA-mediated regulation of PPAR-responsive genes is most likely facilitated by cis9,trans11CLA isomer incorporated in egg yolk. Notably, EFA-CLA activated PPAR more efficiently than nonenriched FA as well as synthetic CLA isomers. We also propose that this regulation, at least in part, can be responsible for the observed reduction in the proliferation of MCF-7 cells treated with EFA-CLA.
Project description:Conjugated Linoleic Acid (CLA) is a collective term for the positional and geometric isomers of linoleic acid. In recent years CLA has been reported to have anti-carcinogenic, anti-obesity, anti-inflammatory and anti-artherogenic effects. The aim of this study was to determine the effects of a natural high-CLA enriched beef source on three primary insulin responsive tissues: adipose, liver and skeletal muscle. ob/ob mice were assigned to one of two diets: a control beef diet, low in c9, t11-CLA (control diet) or a c9, t11-CLA enriched beef diet. After a 28 day feeding trial, RNA was extracted from adipose, liver and skeletal muscle tissue and hybridized to Affymetrix microarrays.
Project description:Iisomer-specific effects of conjugated linoleic (CLA) supplementation on gene expression with particular consideration of the PPAR 2 Pro12Ala SNP in human adipose tissue. Effect of CLA supplementation on genome wide gene expression in adipose tissue biopsies from 5 PPARg2 Ala12Ala and 5 PPARg2 Pro12Pro men were investigated. Subjects underwent four intervention periods (4wk) in a randomized double blind cross-over design receiving f either cis-9, trans-11 CLA, trans-10,cis-12 CLA, 1:1 mixture of both isomers or a reference oil preparation. After each intervention biopsies were taken and whole genome expression microarrays were applied.
Project description:Objective: To quantify changes in adipogenic gene expression in the presence of ritonavir (RTV) or tenofovir (TDF), and determine whether conjugated linoleic acid (CLA) isomers (cis9,trans11 or trans10,cis12) can mitigate detrimental effects of antiretoviral drugs. Methods: Affymetrix Mouse Genome 430 2.0 microarray was used to investigate gene expression in 3T3-L1 adipocytes treated with (1) RTV, TDF or ethanol control, or (2) ritonavir +c9,t11-CLA, ritonavir+t10,c12-CLA or ritonavir+DMSO control. RT-PCR validation of Pparg, Adipoq and Retn was carried out. ELISA and DNA binding ELISA were used to investigate secreted proteins and Pparg binding to its gene response element. Oil Red O staining was used to investigate triglyceride accumulation. Results: No effect was observed for TDF. Expression of 389 genes was altered more than 5-fold in the presence of RTV. Down-regulated genes included Pparg, Adipoq, Retn, Cfd and Cidec. Pparg and Adipoq down-regulation were confirmed by RT-PCR. PPAR-γ binding to its gene response element, adiponectin protein secretion and triglyceride accumulation were decreased by RTV. t10,c12-CLA in the presence of RTV decreased the expression of Ppargand Adipoq in microarray and RT-PCR. c9,t11-CLA increased PPAR-γ binding to its gene response element. Both isomers increased triglyceride storage in the presence of RTV. Conclusion: Ritonavir altered genes involved in adipocyte differentiation, lipid accumulation and glucose metabolism. Down-regulation of Pparg may be mediated by changes in Cepba and regulatory genes Pparg1c and Nr1h3. 3T3-L1 adipocytes were treated with ritonavir (20μM; n=4), tenofovir (1μM; n=4) or vehicle control (ethanol 0.1%; n=4) for 5 days and in a second experiment with ritonavir (20 µM) +c9,t11-CLA (100μM; n=4), ritonavir (20 µM) +t10,c12-CLA (100μM; n=4) or ritonavir+fatty acid vehicle control (DMSO 0.1%; n=4) for 5 days.
Project description:The effects of maternal conjugated linoleic acid (CLA) on embryonic development and hepatic lipid metabolism were investigated in chick embryos. A total of 180 Arbor Acres female broiler breeders (36 wk old) were randomly divided into the following 3 dietary treatment groups: a basic diet (control), a basic diet containing 0.5% CLA (CLA1), and a basic diet containing 1.0% CLA (CLA2). The females were fed for 8 wk, and the eggs from each group were collected and hatched during the last 2 wk. The results showed that the addition of dietary CLA increased the broken egg rate and reduced the fertilization rate and the egg hatchability (P < 0.05). CLA enrichment decreased the polyunsaturated and monounsaturated fatty acids and increased the saturated fatty acids in the yolk sac (P < 0.05). The yolk sac weight, body weight, and body length had a linear decrease with CLA supplementation (P < 0.05). In the developing chick embryo (at E14) and newly hatched chick (D0), the serum triglyceride concentration decreased with maternal CLA supplementation and was accompanied by a reduction in subcutaneous adipose tissue deposition. In addition, maternal CLA supplementation mediated the hepatic lipid metabolism by decreasing the mRNA expression of sterol regulatory element-binding proteins-1c (SREBP-1c), fatty acid synthase and acetyl-CoA carboxylase, and increasing the mRNA expression of adenosine 5'-monophosphate-activated protein kinase ? (AMPK?), peroxisome proliferator-activated receptors ? (PPAR?), liver fatty acid-binding protein, adipose triglyceride lipase and carnitine palmitoyltransferase in embryonic chick livers (P < 0.05). A drop in SREBP-1c protein expression and an increase in the protein expression of p-AMPK? and PPAR? were also observed in the liver of chick embryo (P < 0.05). In conclusion, maternal CLA supplementation regulated the fatty acid composition in the yolk sac, and mediated embryonic chick development and hepatic lipometabolism, and these effects may be related to the AMPK pathway. These findings suggest the potential ability of maternal CLA supplementation to reduce fat deposition in chick embryos.
Project description:Background:Protein quality assessment through the Digestible Indispensable Amino Acid Score requires accurate measurements of true ileal protein and amino acid digestibility, for which a dual isotope technique was recently developed. However, the ileal digestibility of indispensable amino acids (IAA) in humans from high-quality proteins is not well known. Objective:The aim of this study was to intrinsically label hen's egg and meat protein by the use of uniformly 2H-labeled amino acids, and to measure their true ileal indispensable amino acid (IAA) digestibility via the dual isotope method in humans. Design:2H-labeled lyophilized boiled egg white protein, whole boiled egg, and cooked meat were obtained from layer hens (BV-300) administered a uniformly 2H-labeled amino acid mix orally for 35 d with their daily feed. The ileal IAA digestibility of these proteins was determined with reference to digestibility of previously characterized [U-13C]spirulina in a dual tracer method in healthy Indian subjects whose intestinal health was measured by the plasma kynurenine-to-tryptophan (KT) ratio. Results:All subjects had normal KT ratios. The mean ± SD true ileal IAA digestibility of 2H-labeled egg white protein, whole boiled egg, and cooked meat was 86.3% ± 4.6%, 89.4% ± 4.5%, and 92.0% ± 2.8%, respectively. Leucine digestibility correlated with the KT ratio (r = -0.772; P = 0.009). Conclusions:Uniformly 2H-labeled hen's egg and meat protein can be used to measure ileal IAA digestibility by the dual isotope tracer approach in humans. The mean IAA digestibility values for these high-quality proteins in the healthy Indians studied were similar to values obtained in earlier human and animal experiments. Leucine digestibility in these meal matrices correlated with the KT ratio, but this aspect needs further evaluation. This trial was registered at the Clinical Trials Registry of India (http://ctri.nic.in) as CTRI/2018/03/012265.
Project description:Analysis of Helicobacter pylori strain 26695 after 20 minutes of 0.25μg/ml Clarithromycin. Results provide insight into the mechanisms employed by the bacterium that help it adapt to Clarithromycin stress In the study presented here, we compared the gene expression profile between H.pylori without CLA treatment and H.pylori treated with 0.25μg/ml Clarithromycin.
Project description:The aim of this study was to detect miRNAs modified by biactive dietary lipids in a cellular model of enterocytes. The selected model was Caco-2 cells Differentiated Caco-2 cells were incubated with micelles containing the lipids for 24 and 48 hours. The expression leveles of the miRNAs were compared with cells treated with empty micelles. Four control and choelsterol samples, three DHA samples and two CLA samples were analyzed variables: Control (Empty micelles), Cholesterol 250 µM 48h, DHA 200 µM 24h, CLA 200µM 24h Biological replicates Control: Samples 137, 159, 160, 210 Cholesterol: Samples 150, 151, 152, 219 DHA: Samples 141, 142, 143 CLA: Samples 144, 146