RESUMEN
The incidence of liver cancer, the third cause of cancer-associated death, has been growing, worldwide. The increasing trend of liver cancer incidence and mortality indicates the inefficiency of current therapeutic approaches, especially anticancer chemotherapy. Owing to the promising anticancer potential of Thiosemicarbazone (TSC) complexes, this work was conducted to synthesize titanium oxide nanoparticles conjugated with TSC through glutamine functionalization (TiO2@Gln-TSC NPs) and characterize their anticancer mechanism in HepG2 liver cancer cells. Physicochemical analyses of the synthesized particles, including FT-IR, XRD, SEM, TEM, Zeta potential and DLS, and EDS-mapping confirmed the proper synthesis and conjugation of TiO2@Gln-TSC NPs. The synthesized NPs were almost spherical, with a size range of 10-80 nm, a zeta potential of - 57.8 mV, a hydrodynamic size of 127 nm, and without impurities. Investigation of the cytotoxic effect of TiO2@Gln-TSC in HepG2 and HEK293 human normal cells indicated significantly higher toxicity in cancer cells (IC50 = 75 µg/mL) than normal cells (IC50 = 210 µg/mL). Flow cytometry analysis of TiO2@Gln-TSC treated and control cells showed that the population of apoptotic cells considerably increased from 2.8 to 27.3% after treatment with the NPs. Moreover, 34.1% of the TiO2@Gln-TSC treated cells were mainly arrested at the sub-G1 phase of the cell cycle, which was significantly greater than control cells (8.4%). The Hoechst staining assay showed considerable nuclear damage, including chromatin fragmentation and the appearance of apoptotic bodies. This work introduced TiO2@Gln-TSC NPs as a promising anticancer compound that could combat liver cancer cells through apoptosis induction.
RESUMEN
Despite various efforts to produce potent recombinant bio-adhesive proteins for medical purposes, efficient production of a safe and feasible bio-glue is not yet a commercial reality due to the weak properties or low expression levels. Here, a feasible expression system has been developed to produce strong recombinant fusion bioinspired protein using mussel foot protein 3 and 5 (Mfps) along with gas vesicle protein A (GvpA) of Anabaena flos-aquae, and a curli protein CsgA from E. coli, expressed under the control of alcohol oxidase (AOX1) promoter for high-level production in yeast P. pastoris using pPICZα vector. Purified chimeric proteins were first evaluated using western blotting, and their remaining dihydroxyphenylalanine (DOPA) was measured in the modified proteins by NBT assay. We further elucidated the mechanistic properties of obtained adhesive protein assembly in various pH levels based on its different subunits using atomic force microscopy (AFM) when adsorbed onto the mica surface. We found that both combinational structural features of subunits and post-translational changes during expression in yeast host have led to potent adherence due to higher DOPA residues specially in acidic condition and tetrad complex which is higher than that of earlier reports in prokaryotic systems. We believe that our obtained chimeric protein resulted from the fusion of GvpA and CsgA proteins with DOPA-containing Mfp proteins, expressed in the methylotrophic yeast, P. pastoris, not only presents a candidate for future biomedical applications but also provides novel biological clues used for high-performance bioinspired biomaterial designation.
RESUMEN
The aim of this study was to consider the expression of farnesoid X receptor (Fxr), liver X receptor (LXRα) and sirtuin 1 (Sirt1), oxidative stress, inflammation, apoptosis, and the protective role of N-acetylcysteine (NAC) in the liver of rats treated with cadmium (Cd). 30 Wistar rats were divided into 5 groups: G1 (control), G2 (single dose of Cd), G3 (continuous dose of Cd), G4 (single dose of Cd + continuous dose of NAC), and G5 (continuous dose of Cd + continuous dose of NAC). The apoptosis of hepatic cells was measured using the TUNEL assay. Levels of malondialdehyde (MDA), IL-10, TNF-α, and total antioxidant capacity (TAC) were measured by specific kits. The expression of Fxr, LXRα, and Sirt1 genes and ratio of Bax/Bcl2 was considered using RT-PCR. While NAC treatment improved TAC and IL-10 values, it decreased MDA and TNF-α levels in the liver of rats exposed to Cd (P < 0.001). NAC decreased Bax/Bcl2 in the liver of G4 and G5 groups (P < 0.001). Exposure to a continuous dose of Cd decreased Fxr, LXRα, and Sirt1 expression by 36.65- (P < 0.001), 12.52- (P < 0.001) and 11.34-fold (P < 0.001) compared to control, respectively. NAC increased Fxr, LXRα, and Sirt1 expression (P < 0.01) and decreased Cd concentrations in both serum and tissue samples in G4 and G5 groups. Our results suggested that NAC protects liver tissue against Cd toxicity by elevating antioxidant capacity, mitigating oxidative stress, inflammation, apoptosis and up-regulation of FXR, LXR, and SIRT1 genes.