Mitochondria-Mediated Apoptosis of HCC Cells Triggered by Knockdown of Glutamate Dehydrogenase 1: Perspective for Its Inhibition through Quercetin and Permethylated Anigopreissin A

Abstract

Metabolic reprogramming is a hallmark of cancer cells required to ensure high energy needs and the maintenance of redox balance. A relevant metabolic change of cancer cell bioenergetics is the increase in glutamine metabolism. Hepatocellular carcinoma (HCC), one of the most lethal cancer and which requires the continuous development of new therapeutic strategies, shows an up-regulation of human glutamate dehydrogenase 1 (hGDH1). GDH1 function may be relevant in cancer cells (or HCC) to drive the glutamine catabolism from L-glutamate towards the synthesis of α-ketoglutarate (α-KG), thus supplying key tricarboxylic acid cycle (TCA cycle) metabolites. Here, the effects of hGLUD1 gene silencing (siGLUD1) and GDH1 inhibition were evaluated. Our results demonstrate that siGLUD1 in HepG2 cells induces a significant reduction in cell proliferation (58.8% ± 10.63%), a decrease in BCL2 expression levels, mitochondrial mass (75% ± 5.89%), mitochondrial membrane potential (30% ± 7.06%), and a significant increase in mitochondrial superoxide anion (25% ± 6.55%) compared to control/untreated cells. The inhibition strategy leads us to identify two possible inhibitors of hGDH1: quercetin and Permethylated Anigopreissin A (PAA). These findings suggest that hGDH1 could be a potential candidate target to impair the metabolic reprogramming of HCC cells.

PPAR Alpha as a Metabolic Modulator of the Liver: Role in the Pathogenesis of Nonalcoholic Steatohepatitis (NASH)

“…However, other sites within mitochondria can produce ROS [115]. Like peroxisomes, mitochondria contain antioxidant defense systems: superoxide dismutase 1/2 (SOD1/SOD2) enzymes, glutathione/glutaredoxin, thioredoxin/peroxiredoxin systems and low-molecular-weight antioxidants such as CoA, ubiquinol and vitamin C [116].…”

Quercetin, the Ingredient of Xihuang Pills, Inhibits Hepatocellular Carcinoma by Regulating Autophagy and Macrophage Polarization

“…Quercetin inhibits the growth factor-induced migration of HCC cells by inhibiting AKT signaling [38]. Quercetin may also restrain the progress of HCC by inhibiting human glutamate dehydrogenase 1 to regulate mitochondrial function and metabolism [39]. Quercetin inhibits HCC cell migration and invasion, and promotes HCC apoptosis and autophagy by regulating JAK2 and STAT3 pathways [40].…”

Mitochondrial Role in Intrinsic Apoptosis Induced by a New Synthesized Chalcone in Hepatocellular Carcinoma Cells

“…HepG2 cells (2 × 10 4 ) were seeded in a 96-well plate and treated with 2 µM ETTC or DMSO. At different times after ETTC addition, the medium was removed and the activity of caspase 3/7 or 9 was determined by using a Caspase-Glo ® 3/7 or 9 assay kit (Promega), respectively, as previously described [ 33 ]. Luminescence was determined by a GloMax plate reader.…”

“…HepG2 cells were seeded in a 24-well plate at a density of 2 × 10 5 cells/well and treated with 2 µM ETTC or DMSO for 24 h. The production of the mitochondrial superoxide anion was assessed by the MitoSOX™ Red mitochondrial superoxide indicator (MS, Thermo Fisher Scientific), as previously described [ 33 ]. Cells were stained with 5 μM MS for 45 min at 37 °C in the dark, washed with PBS, and analyzed by the Evos Floid Cell Imaging Station (magnification 40×).…”