ABSTRACT
Generation 4 of polyamidoamine dendrimer (G4-PAMAM) has several biological effects due to its tridimensional globular structure, repetitive branched amides, tertiary amines, and amino-terminal subunit groups liked to a common core. G4-PAMAM is cytotoxic due to its positive charges. However, its cytotoxicity could increase in cancer cells due to the excessive intracellular negative charges in these cells. Furthermore, this work reports G4-PAMAM chemical structural characterization using UHPLC-QTOF-MS/MS (LC-MS) by electrospray ionization to measure its population according to its positive charges. Additionally, the antiproliferative effects and intracellular localization were explored in the HMC-1 and K-562 cell lines by confocal microscopy. The LC-MS results show that G4-PAMAM generated multivalent mass spectrum values, and its protonated terminal amino groups produced numerous positive charges, which allowed us to determine its exact mass despite having a high molecular weight. Additionally, G4-PAMAM showed antiproliferative activity in the HMC-1 tumor cell line after 24 h (IC50 = 16.97 µM), 48 h (IC50 = 7.02 µM) and 72 h (IC50 = 5.98 µM) and in the K-562 cell line after 24 h (IC50 = 15.14 µM), 48 h (IC50 = 14.18 µM) and 72 h (IC50 = 9.91 µM). Finally, our results showed that the G4-PAMAM dendrimers were located in the cytoplasm and nucleus in both tumor cell lines studied.
Subject(s)
Dendrimers/pharmacology , Leukemia/drug therapy , Leukemia/metabolism , Nylons/pharmacology , Antineoplastic Agents/analysis , Antineoplastic Agents/pharmacokinetics , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Chromatography, Liquid , Dendrimers/analysis , Dendrimers/pharmacokinetics , Drug Screening Assays, Antitumor/methods , Humans , Inhibitory Concentration 50 , Intracellular Space/drug effects , Intracellular Space/metabolism , K562 Cells , Leukemia/pathology , Nylons/analysis , Nylons/pharmacokinetics , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry , Tissue DistributionABSTRACT
Glutaminase plays an important role in carcinogenesis and cancer cell growth. This biological target is interesting against cancer cells. Therefore, in this work, in silico [docking and molecular dynamics (MD) simulations] and in vitro methods (antiproliferative and LC-MS metabolomics) were employed to assay a hybrid compound derived from glutamine and valproic acid (Gln-VPA), which was compared with 6-diazo-5-oxo-L-norleucine (DON, a glutaminase inhibitor) and VPA (contained in Gln-VPA structure). Docking results from some snapshots retrieved from MD simulations show that glutaminase recognized Gln-VPA and DON. Additionally, Gln-VPA showed antiproliferative effects in HeLa cells and inhibited glutaminase activity. Finally, the LC-MS-based metabolomics studies on HeLa cells treated with either Gln-VPA (IC60 = 8 mM) or DON (IC50 = 3.5 mM) show different metabolomics behaviors, suggesting that they modulate different biological targets of the cell death mechanism. In conclusion, Gln-VPA is capable of interfering with more than one pharmacological target of cancer, making it an interesting drug that can be used to avoid multitherapy of classic anticancer drugs.
Subject(s)
Antineoplastic Agents , Glutamine , Valproic Acid , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Cell Survival/drug effects , Chromatography, Liquid , Glutaminase/antagonists & inhibitors , Glutaminase/chemistry , Glutamine/chemistry , Glutamine/pharmacology , HeLa Cells , Humans , Mass Spectrometry , Metabolome/drug effects , Metabolomics , Models, Molecular , Valproic Acid/chemistry , Valproic Acid/pharmacologyABSTRACT
We present here the results of GHQ-28 items in its spanish version in a sample of 50 university students in examination period. The mean total score was 8.20. We observed a higher mean total score in medical students (9.40) than in students of other careers (7.00), which indicates greater psychopathology in medical students. This fact is corroborated by the higher scores obtained in etch scale.
