ABSTRACT
Growth differentiation factor 11 (GDF11) has been characterized as a key regulator of differentiation in cells that retain stemness features. Recently, it has been reported that GDF11 exerts tumor-suppressive properties in hepatocellular carcinoma cells, decreasing clonogenicity, proliferation, spheroid formation, and cellular function, all associated with a decrement in stemness features, resulting in mesenchymal to epithelial transition and loss of aggressiveness. The aim of the present work was to investigate the mechanism associated with the tumor-suppressive properties displayed by GDF11 in liver cancer cells. Hepatocellular carcinoma-derived cell lines were exposed to GDF11 (50 ng/ml), RNA-seq analysis in Huh7 cell line revealed that GDF11 exerted profound transcriptomic impact, which involved regulation of cholesterol metabolic process, steroid metabolic process as well as key signaling pathways, resembling endoplasmic reticulum-related functions. Cholesterol and triglycerides determination in Huh7 and Hep3B cells treated with GDF11 exhibited a significant decrement in the content of these lipids. The mTOR signaling pathway was downregulated, and this was associated with a reduction in key proteins involved in the mevalonate pathway. In addition, real-time metabolism assessed by Seahorse technology showed abridged glycolysis as well as glycolytic capacity, closely related to an impaired oxygen consumption rate and decrement in adenosine triphosphate production. Finally, transmission electron microscopy revealed mitochondrial abnormalities, such as cristae disarrangement, consistent with metabolic changes. Results provide evidence that GDF11 impairs cancer cell metabolism targeting lipid homeostasis, glycolysis, and mitochondria function and morphology.
Subject(s)
Bone Morphogenetic Proteins/metabolism , Carcinoma, Hepatocellular/metabolism , Growth Differentiation Factors/metabolism , Lipogenesis , Liver Neoplasms/metabolism , Mitochondria/metabolism , Mitochondria/ultrastructure , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Proliferation , Down-Regulation , Glycolysis , Humans , Liver Neoplasms/pathology , Oxygen Consumption , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , TOR Serine-Threonine Kinases/metabolismABSTRACT
INTRODUCTION AND OBJECTIVE: Non-alcoholic fatty liver disease remains as one of the main liver disorders worldwide. It is widely accepted that is the kind of lipid, rather than the amount deposited in the cells that determines cell damage. Cholesterol and saturated free fatty acids are deleterious lipids when accumulated but, in contrast, there are some valuable lipids that could counteract those with harmful properties. Much of this knowledge arises from studies using a single fatty acid, but the effects of a combination of fatty acids, as obtained by diet has been poorly addressed. In the present work, we were focused to figure out the cellular effect of two different mixes of fatty acids, one with high proportion of saturated fatty acids, and another one with high proportion of unsaturated fatty acids (Mediterranean-like) in a cellular model of steatosis. MATERIAL AND METHODS: Primary mouse hepatocytes from animals fed with a western diet (high fat and carbohydrates diet), were treated with both mixes of fatty acids for 24â¯h. RESULTS: Our data clearly show that only the high unsaturated fatty acid mix induced a decrease in triglycerides (47.5%) and cholesterol (59%) content in steatotic hepatocytes mediating cellular protection associated to the decrement of ROS and oxidative damage. The mixture of high saturated fatty acids exhibited no effects, preserving high levels of cholesterol and triglycerides and oxidative damage. In conclusion, our results show that Mediterranean-like mix of fatty acids exerts cellular protection in steatosis by decreasing triglycerides, cholesterol, ROS content and oxidative damage.
Subject(s)
Diet, Mediterranean , Diet, Western , Fatty Acids, Unsaturated/pharmacology , Hepatocytes/drug effects , Lipid Metabolism/drug effects , Liver/drug effects , Non-alcoholic Fatty Liver Disease/drug therapy , Animals , Antigens, Neoplasm/drug effects , Cells, Cultured , Cholesterol/metabolism , Disease Models, Animal , Hepatocytes/metabolism , Hepatocytes/pathology , Liver/metabolism , Liver/pathology , Male , Mice, Inbred BALB C , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/pathology , Oxidative Stress/drug effects , Phospholipases A1/drug effects , Reactive Oxygen Species/metabolism , Triglycerides/metabolismABSTRACT
BACKGROUND/AIMS: To delineate the hypotriglyceridemic effect of conjugated linoleic acid (CLA) in mice, the effect of this fatty acid on lipoprotein lipase (LPL) and apolipoprotein C-III (ApoCIII) mRNA accumulation in muscle, adipose and liver tissue was studied. METHODS: CD-1 mice were housed in groups of 6 and randomized to one of three experimental diets for 3 weeks: SUC: 65% sucrose by weight; CLA: 1% CLA oil (34.4% c9,t11; 35.1% t10,c12 and 4.1% other conjugated isomers) and 65% sucrose, and DEX: 65% dextrose, as a control. RESULTS: LPL mRNA levels in muscle tissue were increased in the DEX group and in the CLA group (240% increase) compared with the SUC group. In contrast, LPL mRNA levels were 81% lower in epididymal adipose tissue from the CLA group compared with the SUC group. There was no effect of dietary treatments on ApoCIII mRNA accumulation in the liver. CONCLUSIONS: These data suggest that dietary CLA may induce partitioning of circulating triglycerides to muscle tissue, preventing their accumulation in adipocytes.
Subject(s)
Apolipoprotein C-III/metabolism , Linoleic Acids, Conjugated/pharmacology , Lipoprotein Lipase/metabolism , RNA, Messenger/metabolism , Triglycerides/metabolism , Adipose Tissue/chemistry , Adipose Tissue/drug effects , Adipose Tissue/metabolism , Animals , Apolipoprotein C-III/drug effects , Apolipoprotein C-III/genetics , Cholesterol/blood , Dietary Sucrose/administration & dosage , Dietary Sucrose/pharmacology , Epididymis/chemistry , Epididymis/drug effects , Epididymis/metabolism , Fatty Acids, Volatile/blood , Gene Expression Regulation, Enzymologic/drug effects , Glucose/administration & dosage , Hypertriglyceridemia/blood , Linoleic Acids, Conjugated/administration & dosage , Lipoprotein Lipase/drug effects , Lipoprotein Lipase/genetics , Liver/chemistry , Liver/drug effects , Liver/metabolism , Male , Mice , Muscle, Skeletal/chemistry , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , RNA, Messenger/drug effects , Random Allocation , Triglycerides/bloodABSTRACT
Lifestyle changes in Alaskan Natives have been related to the increase of cardiovascular disease and metabolic syndrome in the last decades. Variation of the apolipoprotein E (Apo E) genotype may contribute to the diverse response to diet in lipid metabolism and influence the association between fatty acids in plasma and risk factors for cardiovascular disease. The aim of this investigation was to analyze the interaction between Apo E isoforms and plasma fatty acids, influencing phenotypes related to metabolic diseases in Alaskan Natives. A sample of 427 adult Siberian Yupik Alaskan Natives was included. Fasting glucose, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, Apo A1, and Apo B plasma concentrations were measured using reference methods. Concentrations of 13 fatty acids in fasting plasma were analyzed by gas chromatography, and Apo E variants were identified. Analyses of covariance were conducted to identify Apo E isoform and fatty acid main effects and multiplicative interactions. The means for body mass index and age were 26 ± 5.2 and 47 ± 1.5, respectively. Significant main effects were observed for variation in Apo E and different fatty acids influencing Apo B levels, triglycerides, and total cholesterol. Significant interactions were found between Apo E isoform and selected fatty acids influencing total cholesterol, triglycerides, and Apo B concentrations. In summary, Apo E3/3 and 3/4 isoforms had significant interactions with circulating levels of stearic, palmitic, oleic fatty acids, and phenotypes of lipid metabolism in Alaskan Natives.