Cell Line HepG2 Research Articles

A suite of genome-engineered hepatic cells provides novel insights into the spatiotemporal metabolism of apolipoprotein B and apolipoprotein B-containing lipoprotein secretion.

Apolipoprotein B (APOB)-containing very LDL (VLDL) production, secretion, and clearance by hepatocytes is a central determinant of hepatic and circulating lipid levels. Impairment of any of the aforementioned processes is associated with the development of multiple diseases. Despite the discovery of genes and processes that govern hepatic VLDL metabolism, our understanding of the different mechanistic steps involved is far from complete. An impediment to these studies is the lack of tractable hepatocyte-based systems to interrogate and follow APOB in cells, which the current study addresses. To facilitate the cellular study of VLDL metabolism, we generated human hepatic HepG2 and Huh-7 cell lines in which CRISPR/Cas9-based genome engineering was used to introduce the fluorescent protein mNeonGreen into the APOB gene locus. This results in the production of APOB100-mNeon that localizes predominantly to the endoplasmic reticulum (ER) and Golgi by immunofluorescence and electron microscopy imaging. The production and secretion of APOB100-mNeon can be quantitatively followed in medium over time and results in the production of lipoproteins that are taken up via the LDL receptor pathway. Importantly, the production and secretion of APOB-mNeon is sensitive to established pharmacological and physiological treatments and to genetic modifiers known to influence VLDL production in humans. As a showcase, we used HepG2-APOBmNeon cells to interrogate ER-associated degradation of APOB. The use of a dedicated sgRNA library targeting all established membrane-associated ER-resident E3 ubiquitin ligases led to the identification of SYNV1 as the E3 responsible for the degradation of poorly lipidated APOB in HepG2 cells. In summary, the engineered cells reported here allow the study of hepatic VLDL assembly and secretion and facilitate spatiotemporal interrogation induced by pharmacologic and genetic perturbations.

Pomegranate extract-loaded surfactant-free zein nanoparticles as a promising green approach for hepatic cancer: optimization and in vitro cytotoxicity

BackgroundHepatic cancer endures a major health scourge as the consequence of a high incidence of > 1 million cases by 2025. Plant-based products are typically effective in ameliorating health conditions. Pomegranate peel extract (PE) with its high polyphenolic content has anticancer effects against different types of cancer. Herein, we aimed to maximize the PE chemotherapeutic efficacy by loading it in a suitable delivery system to overcome the limitations of PE, to control its release and to achieve liver targeting.MethodA nanoprecipitation procedure was adopted to incorporate PE into biodegradable and biocompatible natural polymeric zein (ZN)-based nanoparticles (NPs) (PE-ZN NPs). A full factorial design (22 × 31) was developed to study the effects of the formulation variables, namely pH of dispersion, PE-to-ZN ratio and surfactant concentration.ResultsThe optimization revealed a surfactant-free stable PE-ZN NPs formula with a small particle size of 99.5 ± 6.43 nm, high PE encapsulation efficiency % of 99.31% ± 3.64 (w/w) and controlled release of PE over 24 h.ConclusionMoreover, the cytotoxicity of the optimum formula against hepatic cancer HepG2 cell lines was assessed and attained about a 2.5-fold reduction in the inhibitory concentration (IC50) values compared to the free PE affording a promising green platform to combat hepatic cancer.Graphical abstract

Multi-omics reveals new links between Fructosamine-3-Kinase (FN3K) and core metabolic pathways

Fructosamine-3-kinases (FN3Ks) are a conserved family of repair enzymes that phosphorylate reactive sugars attached to lysine residues in peptides and proteins. Although FN3Ks are present across the Tree of Life and share detectable sequence similarity to eukaryotic protein kinases, the biological processes regulated by these kinases are largely unknown. To address this knowledge gap, we leveraged the FN3K CRISPR Knock-Out (KO) HepG2 cell line alongside an integrative multi-omics study combining transcriptomics, metabolomics, and interactomics to place these enzymes in a pathway context. The integrative analyses revealed the enrichment of pathways related to oxidative stress response, lipid biosynthesis (cholesterol and fatty acids), and carbon and co-factor metabolism. Moreover, enrichment of nicotinamide adenine dinucleotide (NAD) binding proteins and localization of human FN3K (HsFN3K) to mitochondria suggests potential links between FN3K and NAD-mediated energy metabolism and redox balance. We report specific binding of HsFN3K to NAD compounds in a metal and concentration-dependent manner and provide insight into their binding mode using modeling and experimental site-directed mutagenesis. Our studies provide a framework for targeting these understudied kinases in diabetic complications and metabolic disorders where redox balance and NAD-dependent metabolic processes are altered.

Red Sea Sponge Callyspongia siphonella Extract Induced Growth Inhibition and Apoptosis in Breast MCF-7 and Hepatic HepG-2 Cancer Cell Lines in 2D and 3D Cell Cultures.

The increasing incidence of cancer diseases necessitates the urgent exploration of new bioactive compounds. One of the trends in drug discovery is marine sponges which is gaining significant support due to the abundant production of natural pharmaceutical compounds obtained from marine ecosystems. This study evaluates the anticancer properties of an organic extract from the Red Sea sponge Callyspongia siphonella (C. siphonella) on HepG-2 and MCF-7 cancer cell lines. C. siphonella was collected, freeze-dried, and extracted using a methanol-dichloromethane mixture. The extract was analyzed via Liquid Chromatography-Mass Spectrometry. Cytotoxic effects were assessed through cell viability assays, apoptosis detection, cell cycle analysis, mitochondrial membrane potential assays, scratch-wound healing assays, and 3D cell culture assays. Fifteen compounds were identified in the C. siphonella extract. The extract showed moderate cytotoxicity against MCF-7 and HepG-2 cells, with IC50 values of 35.6 ± 6.9 μg/mL and 64.4 ± 8 μg/mL, respectively, after 48 hours of treatment. It induced cell cycle arrest at the G2/M phase in MCF-7 cells and the S phase in HepG-2 cells. Apoptosis increased significantly in both cell lines, accompanied by reduced mitochondrial membrane potential. The extract inhibited cell migration, with notable reductions after 24 and 48 hours. In 3D cell cultures, the extract had IC50 values of 5.1 ± 2 μg/mL for MCF-7 and 166.4 ± 27 μg/mL for HepG-2 after 7 days of treatment, showing greater potency in MCF-7 spheres compared to HepG-2 spheres. The anticancer activity is attributed to the bioactive compounds. The C. siphonella extract's ability to induce apoptosis, disrupt mitochondrial membrane potential, and arrest the cell cycle highlights its potential as a novel anticancer agent. Additional research is required to investigate the underlying mechanism by which this extract functions as a highly effective anticancer agent.

1H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies

The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.

Design, synthesis, and biological antitumor evaluation of tetrahydroisoquinoline derivatives

Cancer, as a public health issue, is the leading cause of death worldwide. Tetrahydroisoquinoline derivatives have effective biological activities and can be used as potential therapeutic agents for antitumor drugs. In this work, we designed and synthesized a series of novel tetrahydroisoquinoline compounds and evaluated their antitumor activity in vitro on several representative human cancer cell lines. The results showed that the vast majority of compounds showed good inhibitory activities against the cancer cell lines of HCT116, MDA-MB-231, HepG2, and A375.

Enhancing the effectiveness of paclitaxel with stigmasterol bioactive compounds for cancer treatment

This study examines the anticancer activity of stigmasterol bioactive compounds combined with paclitaxel. Drug resistance and high toxicity in chemotherapy present a challenge in treating cancer. However, no study has reported the combination therapy of SS with PTX up to now. We evaluated the potential effects of stigmasterol (SS) combined with Paclitaxel (PTX) on cell viability and apoptosis in liver cancer (HepG2) and normal liver (AML 12) cells. The SS showed the highest cytotoxic effects on HepG2 cells (IC50= 117.75 µg/ml) compared to PTX (IC50= 294.59 µg/ml); however, SS and PTX did not affect normal cells, and SS caused higher apoptotic activity in the sub-G1 phase in HepG2 cells after being treated with SS alone compared to other treatments. In combination therapy for both cells, PTX: SS significantly increased cell viability (IC50 of 464.44 and 1056.44 µg/ml for HepG2 and AML 12, respectively) that differed from the drug alone and the compound, confirming that the PTX combined with SS did not enhance cytotoxic performance against HepG2 cell lines. However, this study found that PTX: SS could reduce the side effects arising from the treatment with PTX. This research indicates that SS is a potential anticancer agent for liver cancer. Furthermore, the current study introduces a novel synergistic therapy that can target various cell lines or enhance the concentration of both SS and PTX to investigate the delivery technologies of both compounds.

Nanoarchitectonics of an acetogenin-enriched nanosystem mediated by an aqueous extract of Annona cherimola Mill with anti-inflammatory and proapoptotic activity against HepG2 cell line

For the first time, this study shows the nanoarchitectonic process to obtain an acetogenin-enriched nanosystem (AuNPs-Ac) using an aqueous extract from Annona cherimola Mill (ACM) composed of gold nanoparticles embedded in an organic matrix that acts as stabilizing agent and presents anti-inflammatory activity and cytotoxical effect against HepG2 cell line, promoting apoptosis. The synthesis of AuNPs-Ac was confirmed by x-ray diffraction analysis, showing metallic gold as the only phase, and the scanning transmission microscope showed an organic cap covering the AuNPs-Ac. Fourier-transformed infrared suggests that the organic cap comprises a combination of different annonaceous acetogenins, alkaloids, and phenols by the presence of bands corresponding to aromatic rings and hydroxyl groups. High-Performance Liquid Chromatography has demonstrated the presence of annonacin, a potent acetogenin, in the extract of ACM. An in vitro anti-inflammatory activity of the extract of ACM and the AuNPs-Ac was performed using the albumin denaturation method, showing a nonlinear response, which is better than sodium diclofenac salt in a wide range of concentrations that goes from 200 to 400 μg ml−1 with both samples. The viability assay was studied using trypan blue, treating IMR90 and HepG2 at different concentrations of AuNPs-Ac. The results defined a median lethal dose of 800 μg ml−1 against HepG2 through apoptosis according to the ratio of caspase-cleaved 9/alpha-tubulin evaluated. It was also demonstrated that the nanosystem presents a higher cytotoxic effect on the HepG2 cell line than in IMR90, suggesting a targeted mechanism. In addition, the nanosystem performs better than using only the extract of ACM in the anti-inflammatory or antiproliferative test, attributed to their higher surface area.

Cephaliverins A-D, four new lignans from Cephalotaxus oliveri Mast.

Four new lignans named cephaliverins A–D (1–4), along with seven known analogues (5–11), were isolated from Cephalotaxus oliveri Mast. Their structures were elucidated on the basis of HR-ESI-MS and NMR analyses, and their absolute configurations were determined by ECD comparison. Cephaliverin A (1), herpetotriol (5) and hedyotol A (6) exhibited moderate antitumor activity against HepG2 and A549 cell lines.

Pyridine-based glycosides bearing 1,2,4-triazole and their 1,3,4-oxadiazole analogues as potential EGFR and CDK-2 inhibitors: Design, synthesis, antiproliferative activity and in silico studies

The increasing toxicity of recently developed anticancer medications is seen as a serious problem that stems from the variety of targets involved; as a result, more research on these phenomena is necessary. In the present work, a novel class of pyridine-based glycosides with a 1,2,4-triazole core and their 1,3,4-oxadiazolyl counterparts with distinct sugar segments are designed and synthesized. The afforded products were studied for their cytotoxicity behavior against a number of cell lines of human cancer. Pyridine-based glycosides 14 and 21 revealed excellent anticancer activity against HepG-2 and MCF-7 cell lines (IC50 = 10.6, 12.8 μM against HepG-2 and 8.3, 9.8 μM against MCF-7, respectively). The inhibitory investigation of the latter promising candidates was further examined in against CDK-2/cyclin A2 kinases and EGFR. The 1,2,4-triazole glycoside-based pyridine system and its oxadiazolyl analog were approximately equipotent with the reference erlotinib in the inhibitory activity against EGFR. On the other hand, the pyridine-1,3,4-oxadiazole product 21 explored a higher potency and sensitivity (half the potency of roscovitine) against CDK-2/cyclin A2 than its pyridine-1,2,4-triazole analogue 14. In an effort to understand its mode of action, it was also investigated how oxadiazolyl-glycoside 21 affected the induction of apoptosis and cell cycle arrest. Furthermore, it increased the MCF-7 cells' p53 and, Bax levels. To further obtain a clear justification and acquire knowledge of the binding nature and affinities of the potent compounds and their targeted enzymes, a molecular docking investigation was simulated. This promotes the deployment of the compounds as effective leads for future investigations in anticancer research.

Targeted pH-responsive delivery of rosmarinic acid via phenylboronic acid functionalized mesoporous silica nanoparticles for liver and lung cancer therapy

Currently, chemotherapy is one of the most practiced approaches for the treatment of cancers. However, existing chemotherapeutic drugs have poor aqueous solubility, poor selectivity, higher systematic toxicity, and poor target accumulation. In this study, we designed and synthesized a boronic acid/ester-based pH-responsive nano-valve that specifically targets the microenvironment in cancer cells. The nano-valve comprises phenylboronic acid-coated mesoporous silica nanoparticles (B-MSN) loaded with polyphenolic compound Rosmarinic acid (ROS-B-MSN). The nano-valve was further coated with lignin (LIG) to achieve our desired LIG-ROS-BMSN nano-valve for targeted chemotherapy against Hep-G2 and NCI-H460 cell lines. The structure and properties of NPs were characterized by Fourier-transformed infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM) in combination with EDX, and Dynamic light scattering (DLS). The outcomes revealed that the designed LIG-ROS-BMSN were in the nanorange (144.1 ± 0.70 nm), had negative Zeta potential (-15.7 ± 0.46 mV) and had a nearly spherical morphology. In vitro, drug release investigations showed a controlled pH-dependent release profile under mild acidic conditions that could enhance the targeted chemotherapeutic response against cancer in mild acidic environments. The obtained LIG-ROS-BMSN nano valve achieved significantly lower IC50 values of (1.70 ± 0.01 μg/mL and 3.25 ± 0.14 μg/mL) against Hep-G2 and NCI-H460 cell lines as compared to ROS alone, which was (14.0 ± 0.7 μg/mL and 29.10 ± 0.25 μg/mL), respectively. The cellular morphology before and after treatment was further confirmed via inverted microscopy. The outcomes of the current study imply that our designed LIG-ROS-BMSN nanovalve is a potential carrier for cancer chemotherapeutics.

Characterization of antibiofilm compound from marine sponge Stylissa carteri.

The fouling phenomenon grabbed global attention and caused huge economic losses specifically in marine-related industries. Sessile behavior exposed the sponge to the risk of fouling. However, their bodies remained free from foulers, which were attributed to the chemical defense system. The objectives of this study were to determine the antibiofilm activity of the marine sponge, Stylissa carteri, and to characterize the isolated compound involved. The antibiofilm activity of S. carteri methanolic crude extract (MCE) and fractions was tested against biofilm-producing bacteria, Pseudomonas aeruginosa, using two different modes of crystal violet biofilm assays: preventive and detachment. Besides that, the disc-diffusion test was conducted to screen the antibacterial activity against gram-positive and gram-negative bacteria while a cytotoxicity assay was conducted on the HepG2 cell line. Bioassay-guided fractionation was carried out using vacuum liquid chromatography (VLC) and solid phase extraction using a C18 Sep-Pak Cartridge. The crystal compound was isolated and characterized through thin-layer chromatography (TLC), Fourier transform infrared (FTIR) spectroscopy, liquid chromatography-mass spectrometry (LCMS), and nuclear magnetic resonance (NMR) spectroscopy. The S. carteri MCE showed a promising result with a half-maximal inhibitory concentration (IC50) of 20.22μg/mL in the preventive assay, while no IC50 was determined in the detachment assay since all inhibitions < 50%. The S. carteri MCE exhibited broad-spectrum antibacterial activity and displayed a non-cytotoxic effect. Fraction 4 from MCE of S. carteri (IC50 = 2.40μg/mL) reduced the biofilm in the preventive assay at all concentrations and exhibited no antibacterial activity indicating the independence of antibiofilm from antibacterial properties. Based on the data obtained, an alkaloid named debromohymenialdisine (DBH) was identified from Fraction 4 of S. carteri MCE. In conclusion, S. carteri was able to reduce the establishment of the biofilm formed by P. aeruginosa and could serve as a prominent source of natural antifouling agents.

Tartaric acid ameliorates experimental non-alcoholic fatty liver disease by activating the AMP-activated protein kinase signaling pathway

Tartaric acid (TA) has been shown beneficial effects on blood pressure and lipid levels. However, its effect on non-alcoholic fatty liver disease (NAFLD) remains unknown. This study aimed to investigate the role of TA in experimental NAFLD. Mice were fed a Western diet for 8 weeks, followed by administration of TA or a vehicle for an additional 12 weeks while continuing on the Western diet. Blood biochemistry including transaminases and glucose tolerance test and liver tissue RNA sequencing (RNA-seq), lipid content, and histology were investigated. The HepG2 cell line was used to explore the mechanism by which TA regulates lipid metabolism. We found that TA significantly improved weight gain, insulin resistance, hepatic steatosis, inflammation and fibrosis in Western diet-fed mice. By comparing gene expression differences, we found that TA affects pathways related to lipid metabolism, inflammatory response, and fibrosis. Furthermore, TA effectively reduced oleic acid-induced lipid accumulation in HepG2 cells and downregulated the genes associated with fatty acid synthesis, which were enriched in the AMP-activated protein kinase (AMPK) signaling pathway. TA also enhanced the phosphorylation of AMPK which could be reverted by the AMPK inhibitor Compound C in HepG2 cells. Our study suggests that TA improves experimental NAFLD by activating the AMPK signaling pathway. These findings indicate that TA may serve as a potential therapy for the human NAFLD.

Assessment of antibacterial, anti-inflammatory, and anti-cancer activities of Melia azedarach L. leaf extract

The purpose of the present study was to evaluate the antibacterial, anti-inflammatory, and cytotoxic properties of Melia azedarach leaf. The phytoconstituents found in the leaves were extracted with four different solvent systems based on their polarity index. Antimicrobial activity of selected plant extracts was evaluated using disc diffusion and micro-broth dilution methods against four different pathogenic bacterial strains. The ethanolic extract of M. azedarach leaf showed a significant inhibitory effect on the growth of Escherichia coli (15.07 mm) and Staphylococcus aureus (18.23 mm) (P &lt; 0.005), followed by Bacillus subtilis and Salmonella typhi. Further, the mechanism of action was confirmed by live/dead cells analysis, which revealed the death of E. coli and S. aureus upon treatment with ethanolic extract. In vivo anti-inflammatory test conducted using carrageenan-induced rat paw edema model revealed that the 300 mg/kg ethanolic extract exhibited a significant anti-inflammatory activity of 51.78% compared with that of standard drug diclofenac (P &lt; 0.001). Further, the cytotoxicity of ethanolic extract against human hepatocarcinoma cell lines (HepG2) was evaluated by MTT assay, and the findings showed a moderate level of toxicity against the HepG2 cell line with an IC50 value of 540.00 &amp;plusmn; 0.6 &amp;mu;g/mL compared to doxorubicin. HepG2 cells treated with doxorubicin (2 &amp;mu;g/mL) and ethanolic extract showed a 2.33- and 1.35-fold increase in p53 gene expression, respectively. Apoptosis activity was measured in terms of DNA laddering, which indicated the late stage of apoptosis. The results showed that the extract-treated cell lines induced DNA fragmentation, which was found to be a potent anti-cancer mechanism in HepG2 cells. This study corroborated that the leaf extract of M. azedarach is a good source of active phytochemicals, with promising biological activities.

Gamma radiation assisted green synthesis of hesperidin-reduced graphene oxide nanocomposite targeted JNK/SMAD4/MMP2 signaling pathway

In this study, a novel method for the fabrication of hesperidin/reduced graphene oxide nanocomposite (RGOH) with the assistance of gamma rays is reported. The different RGOHs were obtained by varying hesperidin concentrations (25, 50, 100, and 200 wt.%) in graphene oxide (GO) solution. Hesperidin concentrations (25, 50, 100, and 200 wt.%) in graphene oxide (GO) were varied to produce the various RGOHs. Upon irradiation with 80 kGy from γ-Ray, the successful reduction of GO occurred in the presence of hesperidin. The reduction process was confirmed by different characterization techniques such as FTIR, XRD, HRTEM, and Raman Spectroscopy. A cytotoxicity study using the MTT method was performed to evaluate the cytotoxic-anticancer effects of arbitrary RGOH on Wi38, CaCo2, and HepG2 cell lines. The assessment of RGOH’s anti-inflammatory activity, including the monitoring of IL-1B and IL-6 activities as well as NF-kB gene expression was done. In addition, the anti-invasive and antimetastatic properties of RGOH, ICAM, and VCAM were assessed. Additionally, the expression of the MMP2-9 gene was quantified. The assessment of apoptotic activity was conducted by the detection of gene expressions related to BCl2 and P53. The documentation of the JNK/SMAD4/MMP2 signaling pathway was ultimately accomplished. The findings of our study indicate that RGOH therapy has significant inhibitory effects on the JNK/SMAD4/MMP2 pathway. This suggests that it could be a potential therapeutic option for cancer.

Propolis Nanoparticles Combined with Gamma-Irradiation Enhance the Potency of Anticancer Against HepG2 Cell Lines: Invitro Investigation

Propolis Nanoparticles Combined with Gamma-Irradiation Enhance the Potency of Anticancer Against HepG2 Cell Lines: Invitro Investigation

Nutritional values, antioxidant, and cytotoxic activities of selected edible marine macroalgae: a comparative study

AbstractThe objective of this study is to assess the chemical composition, nutritional values, and bioactivities of four macroalgae from the Egyptian Red Sea coasts using standard methods. Of these, three Rhodophyceae species, Digenea simplex (D. simplex), Laurencia papillosa (L. papillosa), and Galaxaura oblongata (G. oblongata), and one Phyaeophyceae species, Turbinaria decurrens (T. decurrens), were selected. The results of proximate and chemical composition analyses based on the algal dry weight (DW) showed that carbohydrate content was the highest, ranging from 32.47 ± 1.03% to 45.5 ± 1.23%. Other algal constituents, including ash, protein, moisture, sulfate, lipid, phenolics, and flavonoids, have contents that depend on the algal species. Besides, HPLC analysis revealed that each algal extract contained varying amounts of nine phenolic acids. Atomic absorption spectrometry, the HPLC-Pico-Tag method, the ion chromatography technique, and GC/MS analyses were used to determine the chemical profiles of the elemental, amino acid, halide, and fatty acid of each algal extract. Bioactivities revealed that both the •DPPH and ABTS assays showed that all the algae studied had a significant ability to scavenge free radicals in a dosage-dependent way. They also had strong selective cytotoxic activity against HEPG-2 and HCT-116 cell lines, but only weak activity against MCF-7 and A549 cell lines. Finally, our findings suggested that the selected algae might be efficiently used as nutraceuticals and functional foods, indicating an increase in their proliferation.

Spectroscopic, biological activity, molecular docking and hydrogen storage studies of tetrazole-thione complexes

Six nano-tetrazole-thione complexes of the type [M(MmtzS)2] (MII = Ni (1); Zn(4); Cd (5) and Hg (6)} and [M2(MmtzS)4] (MII = Pd (2) and Pt (3)} were synthesized by treating by the reaction of one equivalent mole of inorganic salts {NiCl2·6H2O; Na2PdCl4; K2PtCl4; ZnCl2; CdCl2 and HgCl2} with two equivalent moles of 1-methyl-1,2,3,4-tetrazole-5-thiol (HMmtzS). The synthesized compounds were obtained through investigation by several physicochemical and spectroscopic techniques. XRD and SEM investigations indicate the crystalline and morphological structural studies of the complexes. Additionally, the MmtzSH ligand and their complexes were tested as antimicrobial agents against four pathogenic bacteria species (Escherichia coli, Klebsiella pneumonia, staphylococcus epidermidis, and Staphylococcus aureus), the results showed that the Ni(II) complex displayed the uppermost activity with the region inhibition within 19 mm(activity index = 72%) to 27 mm (activity index= 96 %) compared to the standard antibacterial drug and other complexes. The cytotoxic effect study was applied to colon carcinoma (CaCo2), and hepatoblastoma (HepG2) cell lines, using the MMT method for complexes (1–4). The Ni(II) complex has the highest activity among all tested compounds. In addition, the complexes 4–6 were chosen for hydrogen storage ability. The findings demonstrate the hydrogen-storing capacity of all tested complexes, with the zinc complex demonstrating the highest capacity at 40 bar and 173 K with 0.99 wt.%.

Targeted delivery of interleukin-12 plasmid into HepG2 cells through folic acid conjugated graphene oxide nanocarrier.

Successful gene therapy relies on carriers to transfer genetic materials with high efficiency and low toxicity in a targeted manner. To enhance targeted cell binding and uptake, we developed and synthesized a new gene delivery vector based on graphene oxide (GO) modified by branched polyethyleneimine (BPEI) and folic acid (FA). The GO-PEI-FA nanocarriers exhibit lower toxicity compared to unmodified PEI, as well as having the potential to efficiently condense and protect pDNA. Interestingly, increasing the polymer content in the polyplex formulation improved plasmid transfer ability. Substituting graphene oxide for PEI at an N/P ratio of 10 in the HepG2 and THP1 cell lines improved hIL-12 expression by up to approximately eightfold compared to simple PEI, which is twice as high as GO-PEI-FA in Hek293 at the same N/P ratio. Therefore, the GO-PEI-FA described in this study may serve as a targeting nanocarrier for the delivery of the hIL-12 plasmid into cells overexpressing folic acid receptors, such as those found in hepatocellular carcinoma.

Integrated in silico and in vitro discovery of a new anticancer thiadiazole analog targeting VEGFR-2

In this work, a new thiadiazole analogue (TDA) was designed with the intention of incorporating key features of Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) inhibitors, and then was examined computationally and experimentally. In vitro (cytotoxicity on MCF-7, HepG2 and WI-38 cell lines, in vitro VEGFR-2 assay, cell cycle analysis, apoptosis assays, RT-PCR, and migration studies), in addition to in silico (molecular docking, ADMET, MD simulations and DFT studies) were conducted. Prior to synthesis, docking and ADMET studies yielded predictions indicating favorable activity and drug likeness. After synthesis, the results demonstrated promising inhibitory activity of TDA on MCF-7 and HepG2 cancer cell lines. These results were notably close to those of sorafenib. The in vitro VEGFR-2 assay revealed significant inhibition, with an IC50 value of 0.283 µM. Selectivity studies indicated a favorable safety profile, with reduced toxicity towards normal lung cells (WI-38). Cell cycle analysis of TDA revealed a significant arrest in the G0-G1 phase, compared to the control group. Apoptosis assays demonstrated a considerable increase in programmed cell death, particularly in the early stages as opposed to the control group. Additionally, there was a notable increase in the expression of the pro-apoptotic gene Bax and a downregulation of the anti-apoptotic gene Bcl2 in MCF-7 cells treated with TDA. TDA also exhibited an anti-angiogenic potential through decreasing the abilities of the MCF-7 cells to migrate and heal. Computationally, various MD simulations studies indicated stable interactions between TDA and VEGFR-2 over 200 ns on both mechanical and energetic levels. DFT studies confirmed reactivity and stability of TDA. These comprehensive findings emphasize TDA’s potential as a multi-targeted candidate for further development in cancer therapy. The promising inhibitory activity, favorable safety profile, and mechanistic insights highlight TDA as a noteworthy contender through further in vivo studies and modifications.