GDF11 restricts aberrant lipogenesis and changes in mitochondrial structure and function in human hepatocellular carcinoma cells.

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.

PPARα via HNF4α regulates the expression of genes encoding hepatic amino acid catabolizing enzymes to maintain metabolic homeostasis.

The liver is the main organ involved in the metabolism of amino acids (AA), which are oxidized by amino acid catabolizing enzymes (AACE). Peroxisome proliferator-activated receptor-α (PPARα) stimulates fatty acid ß-oxidation, and there is evidence that it can modulate hepatic AA oxidation during the transition of energy fuels. To understand the role and mechanism of PPARα's regulation of AA catabolism, the metabolic and molecular adaptations of Ppara-null mice were studied. The role of PPARα on AA metabolism was examined by in vitro and in vivo studies. In wild-type and Ppara-null mice, fed increasing concentrations of the dietary protein/carbohydrate ratio, we measured metabolic parameters, and livers were analyzed by microarray analysis, histology and Western blot. Functional enrichment analysis, EMSA and gene reporter assays were performed. Ppara-null mice presented increased expression of AACE in liver affecting AA, lipid and carbohydrate metabolism. Ppara-null mice had increased glucagon/insulin ratio (7.2-fold), higher serum urea (73.1 %), lower body protein content (19.7 %) and decreased several serum AA in response to a high-protein/low-carbohydrate diet. A functional network of differentially expressed genes, suggested that changes in the expression of AACE were regulated by an interrelationship between PPARα and HNF4α. Our data indicated that the expression of AACE is down-regulated through PPARα by attenuating HNF4α transcriptional activity as observed in the serine dehydratase gene promoter. PPARα via HNF4α maintains body protein metabolic homeostasis by down-regulating genes involved in amino acid catabolism for preserving body nitrogen.

Factores dietéticos y ruptura prematura de membranas: efecto de la vitamina C en la degradación de colágena en el corioamnios / Dietetic factors and premature membranes rupture: vitamin C effect on collagen degradation at amniochorion

Diferentes estudios han mostrado asociación entre la disponibilidad de vitamina C ( vit C) y el desarrollo de ruptura prematura de membranas (RPM). Sin embargo, no ha analizado el papel que desempeña la vit C en el metabolismo de la colágena en el tejido corioamniótico. En este trabajo se analizó el efecto de modulación de diferentes concentraciones de vit C en células en cultivo derivativas de amnios humano. Se utilizaron concentraciones de vit C de manera de cubrir el rango fisiológico (29.0 µg/mL). Luego de ser estimuladas, los medios de las células fueron analizados para actividad enzimática de metaloproteasas de matriz extracelular (MMP) y se cuantificó la cantidad relativa de MMP-1, MMP-2 y MMP-9 mediante inmunotransferencia, utilizando anticuerpos policlonales monoespecíficos. Tanto la actividad como la proteína en los medios de las células amnióticas, disminuyó de manera directa a la concentración de vit C, de manera que a las concentraciones más altas probadas (100 µg/mL) se obtuvo la menor actividad/cantidad de MMP. Los resultados anteriores aportan un dato hasta ahora no descrito y que permite establecer una conexión directa entre la disponibilidad de vitamina C y el aumento en la degradación de colágeno. De acuerdo a los resultados, a menor disponibilidad de vit C, mayor degradación de colágena, que debería llevar a pérdida de soporte mecánico y eventual ruptura de las membranas fetales