Liver Carcinoma HepG2 Cells Research Articles

Effects of prometryn on cardiac and liver cell lines and mouse tissues

BackgroundPesticides are important biological agents that deter or kill pests. Globally, about 2 million tons of pesticides are being utilized each year which highlights their importance. Previous studies have suggested that high levels of pesticide exposure are associated with increased risk of chronic diseases, including cancers, cardiotoxicity, Parkinson’s disease, diabetes mellitus, neurological deficits, birth defects, and reproductive disorders. Prometryn is a commonly used selective triazine herbicide, which is currently used as an alternative to atrazine‐based products. It is widely applied to agriculture as it has lower acute toxicity than atrazine herbicides, stable chemical properties, and a long period of effectiveness. Although past studies have shown that triazine herbicides have carcinogenic potential in humans, the cytotoxic effects of prometryn on cardiac and hepatic systems are not well investigated or understood. The aim of this study was to determine the effects of prometryn on the heart and liver.MethodsH9c2 cardiac and HepG2 liver carcinoma cells were used to study the in‐vitro effects of prometryn. Cells were treated with varying concentrations of prometryn for 24 h. Cell viability, mitochondrial membrane potential, and reactive oxygen species (ROS) levels were assessed. For animal studies, male mice (10 weeks old) (n=24) were randomized to two treatment groups (control and prometryn). The control group received corn oil while the prometryn treated group were exposed to 185mg/kg b.w. prometryn dissolved in corn oil every 48 hours for 7 days. After a total of 4 doses, mice were euthanized, and the heart and liver tissues were collected. Proteasome and immunoproteasome activity were measured in the heart and liver. Protein expression levels of markers for oxidative stress, apoptosis, and proteasomal degradation such as GSTT2, GSTA1/2, Hsp70, Hsp27, NADPH oxidase, PARP‐1, p53, PSMD12, LMP2, DNP, VU‐1 were detected using immunoblotting. Protein oxidation and polyubiquitination were detected as an indicator of oxidative stress. A paired t‐test was used to assess group differences for protein levels and p values <0.05 were deemed statistically significant.ResultWe found that prometryn had a concentration‐dependent effects on H9c2 and HepG2 cell lines when we assessed cell viability, mitochondria function, and total intracellular reactive oxygen species formation. Increasing the concentration of prometryn increased the ROS formation, decreased cell viability and decreased mitochondrial membrane potential in these cell lines. Furthermore, in animal studies we observed changes in the proteasome and immunoproteasome activities in the liver but not in the heart. Changes in protein expression levels of oxidative stress and proteasomal degradation markers were only detected in the liver.ConclusionThese results indicate that H9c2 and HepG2 cells are sensitive to prometryn in‐vitro. However, prometryn effects on the heart and liver tissue in male mice is significantly different. Overall, our data supports the deduction that prometryn affected mitochondrial function, induced oxidative stress in cells, but also alters the ubiquitin‐proteasome system and increases oxidative stress in mice liver. These results help towards elucidation of the mechanism by which prometryn could cause diseases.

Non‐nutritive sweeteners competitively inhibit P‐glycoprotein in liver

Non‐nutritive sweeteners (NNS) are popular sugar replacements that provide sweet taste with little to no calories and are used in a wide range of products, including foods, beverages, and medications. Although NNS are recognized as safe for the general population at the Acceptable Daily Intake (ADI) by the Food and Drug Administration, the effects of NNS on specific patient populations and on human development are not completely understood. Previously, our lab has shown that in mice, pups exposed to NNS during early development present alterations in liver detoxification and metabolic enzymes. This led us to question whether NNS may disrupt detoxification in the liver, including transmembrane efflux transporters such as P‐glycoprotein (PGP). PGP plays a major role in the distribution of common xenobiotics and drugs, such as antibiotics, antidepressants, and antivirals, by excreting their conjugated metabolites from liver cells to the general circulation (blood and bile). Mutations in PGP, along with administration of pharmacological inhibitors lead to intracellular accumulation of its substrates and toxicity.We hypothesize that two common NNS, acesulfame potassium (AceK) and sucralose (Sucr), inhibit PGP’s efflux function in liver cells and alter PGP expression, suggesting reduced detoxification capacity in the liver.To investigate the effects of NNS on hepatic PGP, we first exposed the HepG2 liver carcinoma cell line to AceK and Sucr and assessed PGP mRNA expression by qPCR. Cells were treated with varying concentrations of AceK, Sucr, or combined NNS (AceK+Sucr) that spanned levels below and above the ADI for each sweetener for either 24h acute exposure or 72h chronic exposure. In parallel, PGP protein levels were assessed by Western blot. 24h combined exposure of AceK and Sucr, at concentrations above the ADI, increased PGP transcript expression, but had no effect at concentrations below or equal to the ADI. Only levels above the ADI of AceK and Sucr treatment at 72h led to increased PGP mRNA levels. Combined NNS treatment for 72h, however, increased PGP transcript levels at lower concentrations than acute exposure. In western blot analysis of protein expression, concentrations of combined NNS below the ADI increased PGP levels, in contrast to the qPCR data.As PGP inhibitor drugs have been shown to similarly affect its expression, we then wished to understand whether NNS impact PGP’s efflux action. Therefore, a cell‐free assay measuring PGP’s ATPase activity was used. Complementarily, in vitro assays of substrate retention and substrate efflux in HepG2 were performed to measure cellular efflux. We found that physiologically relevant NNS concentrations led to significant stimulation of ATPase activity, similar to potent PGP competitive inhibitors such as Verapamil. Similarly, low concentrations of AceK, Sucr, or combined NNS, comparable to 1/8th to 1 times the ADI for each, significantly retain PGP substrate Calcein‐AM intracellularly, or reduce efflux of PGP substrate Rhodamine 123, confirming PGP inhibition by NNS.Our results provide evidence that two common NNS, AceK and Sucr, are both substrates and inhibitors of PGP, and that both acute and chronic exposure to those NNS impacts PGP expression. Most importantly, altered PGP efflux function was observed after exposure of hepatic cells to sub‐ADI concentrations of NNS within expected exposure to common foods and beverages, suggesting risks for NNS consumers when taking medications that utilize the PGP as a main detoxification transporter.

RETRACTED ARTICLE: In vitro antioxidant, anticancer, anti-inflammatory, anti-diabetic and anti-Alzheimer potentials of innovative macroalgae bio-capped silver nanoparticles

The antagonistic side effects of chemical medications led to the search for safe strategies such as biogenic agents. Correspondingly, this study aims to create biogenic, appropriate, auspicious and innovative therapeutic agents like Galaxaura elongata {GE}, Turbinaria ornata {TO} and Enteromorpha flexuosa {EF} macroalgae-based silver nanoparticles (Ag-NPs). The Ag+ reduction and the creation of Ag[GE]-NPs, Ag[TO]-NPs and Ag[EF]-NPs have been validated using UV–visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and zeta potential analysis, and the chemical composition of macroalgae crude extracts was estimated through gas chromatography–mass spectrometry (GC–MS). Further, macroalgae-based Ag-NPs were tested for their free radical scavenging activity DPPH, ABTS, anticancer activity in human liver carcinoma (HepG2) cell line, distinctive inflammation forms and elevated α-amylase. Results showed that the biosynthesized Ag-NPs have unique mechanical and physicochemical characters attributed to their high relative surface area, nanosized dimensions and spherical shape. At dose of 200 µg/mL, the DPPH radical scavenging capacity was maximized with Ag[TO]-NPs (67.26%); however, Ag[EF]-NPs was the most potent as ABTs scavenger (97.74%). Additionally, Ag[GE]-NPs had the maximum proteinase inhibitory action with 59.78%. The 1000 µg/mL of Ag[GE]-NPs, Ag[TO]-NPs and Ag[EF]-NPs revealed significant inhibitions of cell growth of HepG2 resulting in cell viabilities 5.92%, 4.44% and 11.33%, respectively. These findings suggest that macroalgae bio-capped Ag-NPs have magnificent biological potentials for safe biomedical applications.

In vitro anticancer activity of silver nanoparticles phyto-fabricated by Hylocereus undatus peel extracts on human liver carcinoma (HepG2) cell lines

Every year, millions of people die from cancer, and most anticancer medications have a low specificity of action and significant toxicity, resulting in severe side effects. As a result, the discovery of effective anticancer drugs is a top priority. Hence, the focal point of this research is to synthesize the dragon fruit peel aqueous extract fabricated silver nanoparticles (DFPAE-AgNPs) in a facile and green chemistry approach as a new-fangled anticancer agent. The selected primary phytochemicals are qualitatively screened for their presence in dragon fruit peel aqueous extract utilized for the fabrication of AgNPs. The analytical results revealed that DFPAE-AgNPs were crystalline, spherical in shape in the size range of 10–50 nm, and composed of 73.90% weight of a silver element. The IC50 value of the DFPAE-AgNPs is evaluated as 37.98 ± 0.21 µg mL−1 with a HepG2 cell line. Acridine orange- ethidium bromide (AO-EtBr) staining and nuclear acid staining assays demonstrated the dose-dependent (10–50 µg mL−1) apoptotic increase phenomenon of the DFPAE-AgNPs in the HepG2 cell line at 24 h. The current study suggested DFPAE as a vital, reliable, and active phytoconstituents for the fabrication of AgNPs with anticancer potentials in a rapid and eco-friendly way. Further meticulous investigation on the DFPAE-AgNPs behavioral nature is recommended to make use of these as alternatives to synthetic drugs in wider anticancer applications.

Mixed Ni(II) and Co(II) complexes of nalidixic acid drug: Synthesis, characterization, DNA/BSA binding profile and in vitro cytotoxic evaluation against MDA-MB-231 and HepG2 cancer cell lines

In this work, herein we report the synthesis, structural characterization and in vitro cytotoxic evaluation of two mixed Co(II)/Ni(II)-nalidixic acid-bipyridyl complexes (1 and 2). The structural analysis of metal complexes 1 and 2 was carried out by analytical and multispectroscopic techniques (FT-IR, UV-vis, EPR, sXRD). The crystallographic details of complexes 1 and 2 revealed a monoclinic crystal system with P21/c space group. DFT studies of complexes were performed to get electronic structure and localization of HOMO and LUMO electron densities. Hirshfeld surface analysis of metal complexes 1 and 2 was employed to understand the various intermolecular interactions (C–H···O, N-H···H and O–H···O) that define the stability of crystal lattice structures. The comparative interaction studies of complex 1 and complex 2 with DNA/BSA were performed by diverse multispectroscopic and analytical techniques to evaluate their chemotherapeutic potential. The magnitude of the DNA binding propensity and binding mode was verified by calculating Kb, K and Ksv values. Higher binding affinity was observed in case of complex 2via intercalative mode. Furthermore, the cytotoxic assessment of complexes 1 and 2 was examined against MDA-MB-231 (triple negative human breast cancer cell line) and HepG2 (liver carcinoma cell line) employing MTT assay which revealed remarkably effecient and specific cytotoxic activity of complex 2.

Degradation of dye wastewater by using cobalt, copper and nickel Schiff base complexes as catalysts: spectral, molecular modelling, catalytic activity and metal removal from aqueous solution

ABSTRACT Metal complexes of Co(II), Cu(II) and Ni(II) with the Schiff base ligand, 6,6ʹ-(([1,1ʹ-biphenyl]-4,4ʹ diylbis(azaneylylidene))bis(methaneylylidene))bis(2,4-dichlorophenol) (H2L) were synthesised. All synthesised compounds were identified and characterised. The conductivity measurements showed the non-electrolytic nature of all the complexes. The decomposition of all complexes ended with metal oxide. Octahedral geometries were proposed for all complexes. Photocatalytic degradation of methylene blue dye (MB) in the presence and absence of hydrogen peroxide was studied by using Co(II), Cu(II) and Ni(II) complexes as catalysts. Fluorescence studies have shown that the reported compounds can act as photoactive materials. Density functional theory was used to investigate the geometries of the ligand and its complexes. The adsorption of Co2+, Ni2+ and Cu2+ in an aqueous solution on H2L ligand under various conditions was studied. The optical properties of all complexes were studied. Antimicrobial activities of the ligand and its transition metal complexes were studied. The in-vitro cytotoxicity of ligand and its cobalt(II), nickel(II) and copper(II) complexes were evaluated towards the human Liver Carcinoma (HepG2) cell line and compared with standard cisplatin.

Green Synthesis of Purslane Seeds-Selenium Nanoparticles and Evaluate its Cytokines Inhibitor in Lead Acetate Induced Toxicity in Rats

Lead is a heavy metal that produces toxicity in both humans and animals due to oxidative stress. The aim of this study was to synthesize through green channel and evaluate hepatoprotective effect of purslane seeds-selenium nanoparticles (PS-Se NPs) against lead acetate-induced liver toxicity. The PS-Se NPs were made and their particle size and zeta potential were measured. In addition, the IC50 of SP-Se NPs against the HepG-2 liver cancer cell line as well as and LD50 in rats were determined. The PS-Se NPs had a mean particle size of around 54.70 ± 4.55 nm, a negative zeta potential of +17.25 and the observed shape of the nanoparticle was spherical. Also, the IC50 of PS-Se NPs against HepG-2 liver carcinoma cell line is 92.66 μg/mL and the LD50 is 1695 mg/kg b.w. Daily oral administration of PS-Se NPs at concentrations of 33.9 and 84.75 mg/kg b.w. for 30 days to rats treated with lead acetate (0.6 mg/kg b.w.) resulted in significant improvements in ALT, AST, ALP, TNF-α, Bcl-2, P53 and MDA. In contrast, oral administration of PS-Se NPs increased the levels of SOD, GPx and GSH in the liver tissue of lead acetate-treated rats. Furthermore, PS-Se NPs almost normalized these effects in histoarchitecture and ultrasound scanning examination of liver tissue. The PS-Se NPs have protective activity against lead acetate-induced liver tissue injury in rats, according to the biochemical, histological and ultrasound scanning findings.

Evaluation of Phytochemical Contents and In Vitro Antioxidant, Anti-Inflammatory, and Anticancer Activities of Black Rice Leaf (Oryza sativa L.) Extract and Its Fractions.

Black rice leaves (Oryza sativa L.) are a major part of rice straw left in open fields after rice harvest as agricultural waste. In this study, crude ethanolic extract (CEE) and various solvent fractions (hexane (Hex), ethyl acetate (EtOAc), n-butanol (n-BuOH), and aqueous fractions) of black rice leaves were investigated for their bioactive compound contents as well as antioxidant, anti-inflammatory, and anticancer activities. The results demonstrated that among all the fractions, the n-BuOH fraction presented the greatest contents of total phenolics and flavonoids, while anthocyanins were found to be abundant in the n-BuOH and aqueous fractions, which also exhibited powerful antioxidant abilities according to DPPH and ABTS radical-scavenging assays and a reducing power assay. Regarding anti-inflammatory activity, CEE and EtOAc reduced the production of NO and cytokine secretion (PGE2, IL-6, and IL-1β) but displayed less effect on tumor necrosis factor α (TNF-α) release in lipopolysaccharide (LPS)-induced RAW 264.7 cells. They also significantly decreased iNOS and COX-2 protein expression. Additionally, the phenolics-rich ethyl acetate fraction showed the greatest activity against HepG2 liver carcinoma cells, inhibited cell growth, increased the Sub-G1 population, and induced apoptosis via mitochondrion-dependent mechanisms. In conclusion, black rice leaves, a byproduct of rice, exhibited strong antioxidant, anti-inflammatory, and anticancer capacities and might be useful for application in functional foods and the pharmaceutical industry.

Assessment of green and chemically synthesized copper oxide nanoparticles against hepatocellular carcinoma

Copper oxide nanoparticle (CuO-NPs) attracts the attentions of human beings due to unique physicochemical and biological properties. The chemical and green synthesis approaches were used to synthesis of CuO-NPs and plant leaves extract Azadirachta indica (A. indica) was preferred in green synthesis approach. The XRD analysis was used to analyze the monoclinic crystal structure and calculate the average crystallite size in the range of 15 ∼ 16 nm. While, the information about different rotational and vibrational modes attached on the spectrum of CuO-NPs was identified by FTIR. After that the UV–VIS analysis provided the information about the absorbance spectrums in the range of 235 and 220 nm. The MTT assay was perfomed to investigate liver carcinoma (HepG2 cells) interaction and absorbance of CuO-NPs towards mentioned cell lines were recorded and loss in HepG2 cells viability. In an overall assessment, anticancer response of comparative study of CuO-NPs towards liver carcinoma treatment contributes significantly after successful demonstration of essential steps of suggested experimental study. Finally, comparative study of experimental and mathematical modeling of anticancer activity towards normal and liver cancer cell lines were conducted and investigated.

Characterization and Antitumoral Activity of Biohybrids Based on Turmeric and Silver/Silver Chloride Nanoparticles.

The phyto-development of nanomaterials is one of the main challenges for scientists today, as it offers unusual properties and multifunctionality. The originality of our paper lies in the study of new materials based on biomimicking lipid bilayers loaded with chlorophyll, chitosan, and turmeric-generated nano-silver/silver chloride particles. These materials showed a good free radical scavenging capacity between 76.25 and 93.26% (in vitro tested through chemiluminescence method) and a good antimicrobial activity against Enterococcus faecalis bacterium (IZ > 10 mm). The anticancer activity of our developed bio-based materials was investigated against two cancer cell lines (human colorectal adenocarcinoma cells HT-29, and human liver carcinoma cells HepG2) and compared to one healthy cell line (human fibroblast BJ cell line). Cell viability was evaluated for all prepared materials after a 24 h treatment and was used to select the biohybrid with the highest therapeutic index (TI); additionally, the hemolytic activity of the samples was also evaluated. Finally, we investigated the morphological changes induced by the developed materials against the cell lines studied. Biophysical studies on these materials were done by correlating UV–Vis and FTIR absorption spectroscopy, with XRD, SANS, and SAXS methods, and with information provided by microscopic techniques (AFM, SEM/EDS). In conclusion, these “green” developed hybrid systems are an important alternative in cancer treatment, and against health problems associated with drug-resistant infections.

A novel bis(pyrazolyl)methane compound as a potential agent against Gram-positive bacteria

This study was designed to propose alternative therapeutic compounds to fight against bacterial pathogens. Thus, a library of nitrogen-based compounds bis(triazolyl)methane (1T–7T) and bis(pyrazolyl)methane (1P–11P) was synthesised following previously reported methodologies and their antibacterial activity was tested using the collection strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Moreover, the novel compound 2P was fully characterized by IR, UV–Vis and NMR spectroscopy. To evaluate antibacterial activity, minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and minimum biofilm eradication concentrations (MBECs) assays were carried out at different concentrations (2–2000 µg/mL). The MTT assay and Resazurin viability assays were performed in both human liver carcinoma HepG2 and human colorectal adenocarcinoma Caco-2 cell lines at 48 h. Of all the synthesised compounds, 2P had an inhibitory effect on Gram-positive strains, especially against S. aureus. The MIC and MBC of 2P were 62.5 and 2000 µg/mL against S. aureus, and 250 and 2000 µg/mL against E. faecalis, respectively. However, these values were > 2000 µg/mL against E. coli and P. aeruginosa. In addition, the MBICs and MBECs of 2P against S. aureus were 125 and > 2000 µg/mL, respectively, whereas these values were > 2000 µg/mL against E. faecalis, E. coli, and P. aeruginosa. On the other hand, concentrations up to 250 µg/mL of 2P were non-toxic doses for eukaryotic cell cultures. Thus, according to the obtained results, the 2P nitrogen-based compound showed a promising anti-Gram-positive effect (especially against S. aureus) both on planktonic state and biofilm, at non-toxic concentrations.

Brefeldin A delivery nanomicelles in hepatocellular carcinoma therapy: Characterization, cytotoxic evaluation in vitro, and antitumor efficiency in vivo

Hepatocellular carcinoma (HCC) is one of the major cancers with high mortality rate. Traditional drugs used in clinic are usually limited by the drug resistance and side effect and novel agents are still needed. Macrolide brefeldin A (BFA) is a well-known lead compound in cancer chemotherapy, however, with poor solubility and instability. In this study, to overcome these disadvantages, BFA was encapsulated in mixed nanomicelles based on TPGS and F127 copolymers (M-BFA). M-BFA was conferred high solubility, colloidal stability, and capability of sustained release of intact BFA. In vitro, M-BFA markedly inhibited the proliferation, induced G0/G1 phase arrest, and caspase-dependent apoptosis in human liver carcinoma HepG2 cells. Moreover, M-BFA also induced autophagic cell death via Akt/mTOR and ERK pathways. In HepG2 tumor-bearing xenograft mice, indocyanine green (ICG) as a fluorescent probe loaded in M-BFA distributed to the tumor tissue rapidly, prolonged the blood circulation, and improved the tumor accumulation capacity. More importantly, M-BFA (10 mg/kg) dramatically delayed the tumor progression and induced extensive necrosis of the tumor tissues. Taken together, the present work suggests that M-BFA has promising potential in HCC therapy.

Pt-Coated Au Nanoparticle Toxicity Is Preferentially Triggered Via Mitochondrial Nitric Oxide/Reactive Oxygen Species in Human Liver Cancer (HepG2) Cells.

Reactive nitrogen species (RNS) that are formed from the reaction of versatile nitric oxide (NO) with reactive oxygen species (ROS) have been less explored in potential cancer therapy. This may be partly due to the fewer available agents that could induce NO in cells. Here, we report platinum-coated gold nanoparticles (Pt-coated Au NPs; 27 ± 20 nm) as a strong inducer of NO (assessed by live-cell imaging under NO-specific DAR-1 probe labeling and indirectly using a Griess reagent) in human liver carcinoma (HepG2) cells. In addition to NO, this study found a critical role of ROS from mitochondrial sources in the mechanism of toxicity caused by Pt-coated Au NPs. Cotreatment with a thiol-replenishing general antioxidant NAC (N-acetyl cysteine) led to significant amelioration of oxidative stress against NP-induced toxicity. However, NAC did not exhibit as much ameliorative potential against NP-induced oxidative stress as the superoxide radical (O2•–)-scavenging mitochondrial specific antioxidant mito-TEMPO did. The higher protective potential of mito-TEMPO in comparison to NAC reveals mitochondrial ROS as an active mediator of NP-induced toxicity in HepG2 cells. Moreover, the relatively unaltered NP-induced NO concentration under cotreatment of GSH modulators NAC and buthionine sulfoximine (BSO) suggested that NO production due to NP treatment is rather independent of the cellular thiols at least in HepG2 cells. Moreover, toxicity potentiation by exogenous H2O2 again suggested a more direct involvement of ROS/RNS in comparison to the less potentiation of toxicity due to GSH-exhausting BSO. A steeper amelioration in NP-induced NO and ROS and, consequently, cytotoxicity by mito-TEMPO in comparison to NAC reveal a pronounced role of NO and ROS via the mitochondrial pathway in the toxicity of Pt-coated Au NPs in HepG2 cells.

Synthesis, Cytotoxicity, Antioxidant and Antimicrobial Activity of Indole Based Novel Small Molecules

Aim: In this study experiments were carried out to explore antioxidant, antimicrobial, cytotoxic properties of novel indole derivative 1-ethyl-2-phenyl-3-phenylethyl-3-thiophen-2-yl-1Hindole (EPI) together with its effect on glutathione S-transferases (GST) activities in human liver carcinoma (HepG2) cells. Background:: Indoles probably represent one of the most important heterocyclic structures that have been attracting the interest of many scientists in drug discovery. Objective: The present study was carried out to explore antioxidant, antimicrobial, cytotoxic properties of novel indole derivative 1-ethyl-2-phenyl-3-phenylethyl-3-thiophen-2-yl-1H-indole (EPI) and its effect on glutathione S-transferases (GST) activities in human liver carcinoma (HepG2) cells. Materials and Methods: Pd-catalyst Sonogashira coupling reactions, MTT Assay, Antioxidant capacity test, Antimicrobial test, GST enzyme activity test. Results: 1-ethyl-2-phenyl-3-(phenylethynyl)-1H-indole had antioxidant and antimicrobial properties. It displayed significant induction in glutathione S-transferases (GST) enzyme activity in human liver cancer cell lines (HepG2), but cytotoxic effect on all tested cancer cell lines could not be observed. Conclusion:: All of these results showed that 1-ethyl-2-phenyl-3-(phenylethynyl)-1H-indole had antioxidant and antimicrobial properties without cytotoxic effect, which could make it a promising active component with further studies.

Individual and combined hepatocytotoxicity of DDT and cadmium in vitro.

The organochlorine insecticide dichlorodiphenyltrichloroethane (DDT) and heavy metal cadmium (Cd) are widespread environmental pollutants. They are persistent in the environment and can accumulate in organisms. Although the individual toxicity of DDT and Cd has been well documented, their combined toxicity is still not clear. Since liver is their common target, in this study, the individual and combined toxicity of DDT and Cd in human liver carcinoma HepG2 and human normal liver THLE-3 cell lines were investigated. The results showed that DDT and Cd inhibited the viability of HepG2 and THLE-3 cells dose-dependently and altered lysosomal morphology and function. Intracellular reactive oxygen species and lipid peroxidation levels were induced by DDT and Cd treatment. The combined cytotoxicity of DDT and Cd was greater than their individual cytotoxicity, and the interaction between Cd and DDT was additive on the inhibition of cell viability and lysosomal function of HepG2 cells. The interaction was antagonistic on the inhibition of cell viability of THLE-3 cells. These results may facilitate the evaluation of the cumulative risk of pesticides and heavy metal residues in the environment.

Selenite Inhibits Notch Signaling in Cells and Mice.

Selenium is an essential micronutrient with a wide range of biological effects in mammals. The inorganic form of selenium, selenite, is supplemented to relieve individuals with selenium deficiency and to alleviate associated symptoms. Additionally, physiological and supranutritional selenite have shown selectively higher affinity and toxicity towards cancer cells, highlighting their potential to serve as chemotherapeutic agents or adjuvants. At varying doses, selenite extensively regulates cellular signaling and modulates many cellular processes. In this study, we report the identification of Delta–Notch signaling as a previously uncharacterized selenite inhibited target. Our transcriptomic results in selenite treated primary mouse hepatocytes revealed that the transcription of Notch1, Notch2, Hes1, Maml1, Furin and c-Myc were all decreased following selenite treatment. We further showed that selenite can inhibit Notch1 expression in cultured MCF7 breast adenocarcinoma cells and HEPG2 liver carcinoma cells. In mice acutely treated with 2.5 mg/kg selenite via intraperitoneal injection, we found that Notch1 expression was drastically lowered in liver and kidney tissues by 90% and 70%, respectively. Combined, these results support selenite as a novel inhibitor of Notch signaling, and a plausible mechanism of inhibition has been proposed. This discovery highlights the potential value of selenite applied in a pathological context where Notch is a key drug target in diseases such as cancer, fibrosis, and neurodegenerative disorders.

Antioxidant and Anti-proliferative Activities of Smilax Zeylanica Root and Rhizome Extract against Liver Carcinoma Cells

ABSTRACT The antioxidant, cytotoxic, and anti-cancer potential of aqueous root and rhizome extract (ARRE) of Smilax zeylanica was investigated against human liver carcinoma cells. The phytochemical composition was evaluated by standard methods, and confirmed by thin-layer chromatography. Free radical scavenging activity was determined by DPPH assay, cytotoxic activity by brine shrimp lethality assay, and anti-cancer potential was investigated using liver carcinoma HepG2 cells. Phytochemical analysis revealed the presence of saponins, flavonoids, proanthocyanins, tannins, and polyphenols. The total phenolic and flavonoid content varied from 0.28 to 5.26 mg gallic acid equivalent 100 mg−1 and 0.25–50.26 quercetin equivalent 100 mg−1, respectively. There was a linear relationship between the free radical scavenging activity and phytochemical concentrations with an IC50 value of 108 ± 1.2 µg mL−1 compared to the standard (IC50: 2.59 ± 0.02 µg mL−1). The brine shrimp lethality assay showed strong cell killing ability (IC50: 84 ± 0.7 µg mL−1) and the extract induced apoptosis of HepG2 cells (77 ± 1.52%) at 300 µg mL−1, comparable to the positive control (camptothecin).

One-Step Synthesis of Magnetic Nanocomposite with Embedded Biologically Active Substance.

Magnetic nanocomposites based on hydroxyapatite were prepared by a one-step process using the hydrothermal coprecipitation method to sinter iron oxides (Fe3O4 and γ-Fe2O3). The possibility of expanding the proposed technique for the synthesis of magnetic composite with embedded biologically active substance (BAS) of the 2-arylaminopyrimidine group was shown. The composition, morphology, structural features, and magnetic characteristics of the nanocomposites synthesized with and without BAS were studied. The introduction of BAS into the composite synthesis resulted in minor changes in the structural and physical properties. The specificity of the chemical bonds between BAS and the hydroxyapatite-magnetite core was revealed. The kinetics of the BAS release in a solution simulating the stomach environment was studied. The cytotoxicity of (HAP)FexOy and (HAP)FexOy + BAS composites was studied in vitro using the primary culture of human liver carcinoma cells HepG2. The synthesized magnetic composites with BAS have a high potential for use in the biomedical field, for example, as carriers for magnetically controlled drug delivery and materials for bone tissue engineering.

Effects of pretreatment and type of hydrolysis on the composition, antioxidant potential and HepG2 cytotoxicity of bound polyphenols from Tartary buckwheat (Fagopyrum tataricum L. Gaerth) hulls

This study investigated the effects of ultrasound assisted-subcritical water (U-SW), subcritical water (SW), ultrasound (U) and hot water (HW) pretreatments and acid hydrolysis (AH) and alkaline hydrolysis (AlkH) on the phenolic composition, antioxidant potential and cytotoxicity of Tartary buckwheat hull extracts. The Folin Ciocalteu assay and HPLC-MS were used to characterize and quantify phenolics of the extracts. The ABTS, FRAP and TEAC assays were used to measure antioxidant activity and the MTT assay was used to measure cytotoxicity of the extracts in HepG2 human liver cancer cells. Results showed that U-SW gave the best AH yield of phenolics (128.45), followed by SW (85.82) and U (64.70), compared to the control, HW (35.82 mgg−1). The same trend was observed for phenols extracted using AlkH. U-SW had the highest antioxidant activity, followed by SW and U regardless of hydrolytic method used. Cytotoxicity followed a similar trend with U-SW and SW being the most cytotoxic to liver cancer cells, followed by U, with the least being HW. The findings suggested that plant materials such as Tartary buckwheat hulls can be pretreated with U-SW, SW and U prior to hydrolytic recovery of bound polyphenols. Also, AH was more efficient than AlkH for phenol extraction, and gave extracts with higher antioxidant activity and cytotoxicity in HepG2 liver carcinoma cells. This application allows for beneficial usage of agricultural biomass and help diversify income sources and products for industry.

Green and chemically synthesized zinc oxide nanoparticles: effects on in-vitro seedlings and callus cultures of Silybum marianum and evaluation of their antimicrobial and anticancer potential

Zinc oxide nanoparticles (ZnO-NPs) have been produced by physical and chemical methods. Here, the comparative evaluation of both chemically-synthesised ZnO-NPs (C-ZNPs) and in-vitro cultured S. marianum mediated green-synthesised ZnO-NPs (G-ZNPs) were investigated on seed germination frequency, root and shoot growth, callus induction and biochemical profile of medicinally important plant Silybum marianum. Of all the treatments, callus-mediated ZnO-NPs gave optimum results for seed germination (65%), plantlet’s root length (4.3 cm), shoot length (5.3 cm) and fresh and dry weights (220.4 g L−1 and 21.23 g L−1, respectively). Similarly, the accumulation of phenolic (12.3 µg/mg DW) and flavonoid (2.8 µg/mg DW) contents were also enhanced in callus cultures treated with G-ZNPs. We also observed maximum antioxidant activity (99%) in callus cultures treated with G-ZNPs, however, in case of plantlets, these activities were found highest for in-vitro whole plant-mediated ZnO-NPs. Moreover, G-ZNPs also enhanced total protein content (265.32 BSAE/20g FW) in callus cultures. G-ZNPs were further assessed for their effects on several multidrug resistant bacterial strains and human liver carcinoma (HepG2) cells and our findings revealed that callus extracts treated with G-ZNPs show ameliorated antibacterial (highest zone of inhibition (19 mm) against Klebsiella pneumonia) and anticancer (highest cytotoxicity of 64%) activities.