Human Liver Cancer Research Articles

Synthesis of Sulfonamide-Based Schiff Bases as Potent Anticancer Agents: Spectral Analyses, Biological Activity, Molecular Docking, ADME, DFT, and Pharmacophore Modelling Studies.

The current study focuses on the synthesis and characterization of six benzenesulfonamide-based Schiff base derivatives (7-12) with various electron-withdrawing and electron-donating substituents (-F, -CI, -Br, -CH3, and -OCH3) and the assessment of their antiproliferative activities against human lung (A549) and liver (HepG2) cancer cell lines using in vitro and in silico approaches. The structures of the synthesized compounds (7-12) were elucidated by elemental analysis and FT-IR, 1D (1H, 13C, APT, and DEPT-135), and 2D (HMQC and HMBC) NMR spectroscopies. The cytotoxic activities of the targeted compounds were determined at various concentrations against these cancer cell lines for 72 h, using the MTT method. The targeted molecules (7-12) demonstrated remarkable antiproliferative activities, with IC50 values ranging from 6.032-9.533 μM against the A549 cell line and 5.244-9.629 μM against the HepG2 cell line. These compounds showed activities at lower or very similar concentrations to cisplatin against the A549 cell line and at much lower concentrations than cisplatin against the HepG2 cell line. Among them, compounds 10 and 12 were found to be more effective against A549 and HepG2 cells, respectively, than cisplatin. These compounds were analyzed by interacting with the 1BNA, 4HJO, and 4ASD crystal structures in molecular docking studies. The docking score of 4ASD-compound 12 interaction was calculated as -4.045 kcal/mol, 4HJO-compound 10 interaction was calculated as -5.179 kcal/mol and 1BNA-compound 10 interaction was calculated as -8.571 kcal/mol and it was determined that these compounds were theoretically better than Cisplatin. In the present study, ADME data were estimated using the web tool SwissADME. With ADME, it was calculated that the logP value of compounds 7-12 was less than 5, the HBD number was 1, the HBA number was 7 or 8, and the molecular weight was less than 500. Properties such as the electrophilic index and chemical hardness of the designed compounds were examined by density functional theory (DFT) using B3LYP/6-311G**. In conclusion, these compounds have emerged as promising new anti-cancer drug candidates.

Green chemistry approach to synthesize titanium dioxide nanoparticles using Fagonia Cretica extract, novel strategy for developing antimicrobial and antidiabetic therapies

Abstract This groundbreaking study explores the eco-friendly production of titanium dioxide nanoparticles (TiO2 NPs) to investigate their impact on health. TiO2 NPs were synthesized utilizing a plant extract from Fagonia cretica, acting as both stabilizers and reducers. Various techniques, including energy dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), UV-Vis, and X-ray diffraction (XRD), were employed to analyze the synthesized TiO2 NPs. FT-IR revealed functional groups crucial for nanoparticle (NP) formation. SEM confirmed the particle size of synthesized TiO2 NPs, ranging from 20 to 80 nm. XRD analysis highlighted the rutile phase crystalline structure, and EDX determined the elemental composition of TiO2 NPs. These NPs displayed potent antimicrobial properties, proving toxic to bacterial and the fungal strains at 50 µg·mL−1 concentration. Impressively, TiO2 NPs showcased significant antidiabetic effects in adult male albino mice, effectively reducing Streptozotocin-induced hyperglycemia and hypercholesterolemia by the improvement in behavior via random blood glucose, triglyceride, low density lipoproteins, high density lipoprteins, very low density lipoproteins, and GTT pathway at 100 and 200 µL. Furthermore, they exhibited a remarkable impact on human liver cancer cell lines, with a 43.2% reduction in cell viability at 100 µg·mL−1 concentration. In essence, the study highlights TiO2 NPs as a safe, natural therapeutic agent with immense potential in diabetes treatment. The MTT assay was utilized to assess their cytotoxicity and biocompatibility, affirming their promising role in healthcare.

Structure–Activity Relationship Studies of Imidazo[1′,2′:1,6]pyrido[2,3-d]pyrimidine Derivatives to Develop Selective FGFR Inhibitors as Anticancer Agents for FGF19-overexpressed Hepatocellular Carcinoma

The aberrant activation of fibroblast growth factor (FGF) and FGF receptor (FGFR)-mediated signaling pathways are associated with cancer development, including hepatocellular carcinoma (HCC). A novel series of imidazo[1′,2′:1,6]pyrido[2,3-d]pyrimidine, containing an acrylamide covalent warhead, were synthesized as selective FGFR 1-4 inhibitors. Compound 7n was identified as the most potent inhibitor against FGFR1, 2, and 4, with IC50 values of 8/4 nM (FGFR1/2) and 3.8 nM (FGFR4), and the covalent docking analyses suggested that 7n form a covalent adduct with cysteine residue on the hinge or p-loop of FGFR. Compound 7n exhibited a favorable pharmacokinetic profile and significant in vivo antitumor efficacy in human liver cancer xenograft mouse models (xenograft, FGF/FGFR-dependent HCC cells).

Tunable Multi-Enzyme Activities of Platinum Nanoclusters for Enhanced Specificity and Sensitivity in Biosensing

Nanozymes have gained prominence for their utility in biosensing and disease diagnostics. However, challenges arise from complex sample matrices and nonspecific enzyme activities that contribute to false signals. This study introduces multifunctional platinum nanoclusters (Pt NCs) exhibiting peroxidase-like (POD-like), oxidase-like (OXD-like), and laccase-like activities tailored for enhanced biosensing capabilities. By adjusting pH, we optimized the conditions to achieve distinct POD-like and OXD-like responses, thereby reducing background signals and improving detection accuracy. The addition of ATP further amplified the POD-like activity while minimizing interference from OXD-like activity. This combined strategy substantially enhanced biomarker detection selectivity, demonstrated through glucose detection in human serum samples. Moreover, thiol inhibition of laccase-like activity in Pt NCs was leveraged for thiol-based antioxidant assessment, revealing their application in quantifying total antioxidant capacity (TAC) in human liver cancer cells, accounting for 44% of TAC. The Pt NCs demonstrated robust sensitivity and reusability, offering a novel multi-enzyme nanomaterial with potential for precise and interference-free biosensing applications. These findings contribute to the development of advanced nanozyme-based biosensors, addressing specificity regulation challenges and expanding their practical application in biosensing and disease diagnostics.

Construction and Evaluation of Hepatic Targeted Drug Delivery System with Hydroxycamptothecin in Stem Cell-Derived Exosomes.

Hydroxycamptothecin (HCPT) is commonly used in the treatment of liver cancer; however, its low water solubility and poor stability significantly limit its clinical application. In recent years, research on exosomes has deepened considerably. Exosomes possess a unique phospholipid bilayer structure, enabling them to traverse tissue barriers, which provides natural advantages as drug carriers. Nevertheless, delivering exosomes safely and efficiently to target cells remains a major challenge. In this study, we utilized the affinity of the SP94 peptide for human liver cancer cell receptors. HCPT was coated with exosomes in our experimental design, and the exosome membrane was modified with SP94 peptide to facilitate drug delivery to liver cancer cells. Exosomes were purified from bone marrow mesenchymal stem cells, and targeted peptides were attached to their surfaces via post-insertion techniques. Subsequently, HCPT was incorporated into the exosomes through electroporation. Using the HepG2 hepatoma cell line, we evaluated a series of in vitro pharmacodynamics and studied pharmacokinetics and tissue distribution in animal models. The results indicated that ligand-targeted, modified drug-carrying exosomes significantly enhance drug bioavailability, prolong retention time in vivo, and facilitate liver targeting. Moreover, this approach reduces drug nephrotoxicity, enhances anti-tumor efficacy, and lays the groundwork for the development of novel liver cancer-targeting agents.

Selective Metal‐Coordinated Nanodrugs Based on Thiazine Moiety for Anticancer Therapeutics: Spectroscopic, Theoretical, and Molecular Docking Studies

ABSTRACTThe current study involved production, comprehensive structural analysis, and physicochemical characterizing of two distinctive complexes namely, Cu(TBH) and Zn(TBH); TBH donor ligand: N′‐(1‐(3,6‐dihydro‐4‐hydroxy‐2,6‐dioxo‐2H‐1,3‐thiazin‐5‐yl)ethylidene)‐2‐hydroxybenzohydrazide) using a wide range of analytical methods, including elemental analysis, UV–Vis, FT‐IR, and 1HNMR spectrometry, molar conductivity and magnetic susceptibility measurements, and thermal analysis. According to the findings, chelation can occur through O, N, and O donor atoms of monoanionic chelator for generating mono‐nuclear chelates with tetrahedral geometry for Cu (II) and octahedral geometry for Zn (II). The anticarcinogenic ability of TBH chelator and its coordinated compounds against human liver cancer (HepG‐2) was investigated. The Cu(TBH) complex was found to have selective and promising anticancer activity against a liver cancer cell line, with lower IC50 (liver carcinoma cell line) and higher IC50 (normal human cell line) values than other produced compounds and standard drugs. Utilizing DFT computations, the molecular structures of the TBH and its complexes were verified, offering a detailed understanding of their quantum chemical characteristics. A quantitative structure–activity relationship (QSAR) model, which illustrates the association between DFT‐computed descriptors and biological activities pIC50, was also created using multiple linear regression (MLR) on the synthesized compounds as anticancer medicines. Additionally, there are no issues with the produced compounds' oral bioavailability according to (ROF) Lipinski's rule of five. Furthermore, docking studies of the synthesized TBH chelator and its chelates with CDK2 kinase have been performed to validate the biological findings. According to our findings, the novel Cu(TBH) nanodrug exhibits considerable cytotoxic activity, prompting further study into its pharmacological profile and exploring its potential in drug development.

Expression of EGFRvIII and its co‑expression with wild‑type EGFR, or putative cancer stem cell biomarkers CD44 or EpCAM are associated with poorer prognosis in patients with hepatocellular carcinoma.

The aberrant expression of HER family members and cancer stem cells (CSCs) have been associated with tumour progression and resistance to therapy. At present, several HER inhibitors have been approved for the treatment of patients with a range of cancers but not for the treatment of patients with hepatocellular carcinoma (HCC). The present study investigated the co‑expression and prognostic significance of HER family members, type‑III deletion mutant EGFR (EGFRvIII), and the putative CSC biomarkers CD44 and epithelial cell adhesion molecule (EpCAM) in 43 patients with HCC. The relative expression of these biomarkers was determined using immunohistochemistry. At a cut off value of >5% of tumour cells stained for these biomarkers, 35% [wild‑type (wt)EGFR], 58% (HER‑2), 0% (HER‑3), 19% (HER‑4), 26% (EGFRvIII), 40% (CD44) and 33% (EpCAM) of patients were positive. In 23, 14 and 9% of the patients, wtEGFR expression was accompanied by co‑expression with HER‑2, EGFRvIII and HER‑2/EGFRvIII, respectively. EGFRvIII expression, membranous expression of CD44 and co‑expression of wtEGFR/EGFRvIII were associated with poor overall survival (OS). By contrast, cytoplasmic CD44 expression was associated with a longer OS time. The present study also investigated the effect of several agents targeting one or more members of the HER family, other growth factor receptors and cell signalling proteins on the proliferation of HCC cell lines. Among agents targeting one or more members of the HER family, the pan‑HER family blocker afatinib was the most effective, inhibiting the proliferation of three out of seven human liver cancer cell lines (LCCLs), while the CDK inhibitor dinacicilib was the most effective agent, inhibiting the proliferation of all human LCCLs tested. Taken together, the present results suggested that EGFRvIII expression and its co‑expression with wtEGFR or CD44 was of prognostic significance. These results also support further investigations of the therapeutic potential of drugs targeting EGFRvIII and other members of the HER family in patients with HCC.

Modulatory Effects of Chalcone Thio-Derivatives on NF-κB and STAT3 Signaling Pathways in Hepatocellular Carcinoma Cells: A Study on Selected Active Compounds.

Our previous studies demonstrated the modulatory effects of new synthetic thio-chalcone derivatives in dishes on the Nrf2, NF-κB, and STAT3 signaling pathways in colon cancer cells. This study aimed to evaluate the effect of four selected active chalcone thio-derivatives on the NF-κB and STAT3 signaling pathways involved in inflammatory processes and cell proliferation in human liver cancer cells. Cell survival was assessed for cancer (HepG2) and normal (THLE-2) cell lines. Activation of NF-κB and STAT3 signaling pathways and the expression of proteins controlled by these pathways were estimated by Western blot, and qRT-PCR assessed the expression of NF-κB and STAT3 target genes. We also evaluated the impact on the selected kinases responsible for the phosphorylation of the studied transcription factors by MagneticBead-Based Multiplex Immunoassay. Among the thio-derivatives tested, especially derivatives 1 and 5, there was an impact on cell viability, cell cycle, apoptosis, and activation of NF-κB and STAT3 pathways in hepatocellular carcinoma (HCC), which confirms the possibilities of using them in combinatorial molecular targeted therapy of HCC. The tested synthetic thio-chalcones exhibit anticancer activity by initiating proapoptotic processes in HCC while showing low toxicity to non-cancerous cells. These findings confirm the possibility of using chalcone thio-derivatives in molecularly targeted combination therapy for HCC.

5149 In-Vitro Combo Treatment of Human Liver Cancer Cell (Hep-G2) Line With Vitamin-A And RU-486

Abstract Disclosure: P. Ramaraj: None. Our previous in-vitro studies involving vitamins (vitamin-A and vitamin-D3) and steroids (progesterone and RU-486) pointed to a combination of vitamin-A and RU-486 as an efficient combo to decrease human melanoma cell growth. We extended this combo treatment to human liver cancer cell (Hep-G2) line. The aim was to check whether vit-A and RU-486 combo would also decrease human liver cancer cell line growth. Liver cancer (Hep-G2) cells were plated in a 96 well plate overnight. Following day, vit-A and RU-486 were added either alone or in different combinations for 48 hrs. After 48 hrs, cell viability and percentage of cell growth were monitored by the MTT assay. Supernatants were collected for Elisarray and quantitation of cytokine(s) by Elisa. Individual vit-A and RU-486 treatments showed a dose-dependent decrease in cell growth. Though various combinations of vit-A (50, 75, 100 μM) and RU-486 (10, 50 100 μM) also showed a decrease in cell growth, an experimentally significant decrease in cell growth was observed at vit-A 75 μM and RU-486 50 μM combination. This combo resulted in 24% cell growth compared to straight vit-A (75 μM) at 44% cell growth and RU-486 (50 μM) at 35% cell growth. Thus, this combo inhibited liver cancer cell growth also. In the future effect of this combo on normal liver cell line and the cytokine(s) if any, playing a central role in cell growth would be identified by Elisarray and quantitated by Elisa. Presentation: 6/3/2024

5149 In-vitro Combo Treatment of Human Liver Cancer cell (Hep-G2) Line with Vitamin-A and RU-486

Abstract Disclosure: P. Ramaraj: None. Our previous in-vitro studies involving vitamins (vitamin-A and vitamin-D3) and steroids (progesterone and RU-486) pointed to a combination of vitamin-A and RU-486 as an efficient combo to decrease human melanoma cell growth. We extended this combo treatment to human liver cancer cell (Hep-G2) line. The aim was to check whether vit-A and RU-486 combo would also decrease human liver cancer cell line growth. Liver cancer (Hep-G2) cells were plated in a 96 well plate overnight. Following day, vit-A and RU-486 were added either alone or in different combinations for 48 hrs. After 48 hrs, cell viability and percentage of cell growth were monitored by the MTT assay. Supernatants were collected for Elisarray and quantitation of cytokine(s) by Elisa. Individual vit-A and RU-486 treatments showed a dose-dependent decrease in cell growth. Though various combinations of vit-A (50, 75, 100 μM) and RU-486 (10, 50 100 μM) also showed a decrease in cell growth, an experimentally significant decrease in cell growth was observed at vit-A 75 μM and RU-486 50 μM combination. This combo resulted in 24% cell growth compared to straight vit-A (75 μM) at 44% cell growth and RU-486 (50 μM) at 35% cell growth. Thus, this combo inhibited liver cancer cell growth also. In the future effect of this combo on normal liver cell line and the cytokine(s) if any, playing a central role in cell growth would be identified by Elisarray and quantitated by Elisa. Presentation: 6/3/2024

Chemical profiling, cytotoxicity and mode of action of sea urchin Echinometra mathaei exoskeleton extract in HepG2 cancer cell line

Marine organisms contain several secondary metabolites with potent biological and pharma­cological activities. This research aimed to study the cytotoxicity and mechanism of action of methanol (MeOH) extract from the exoskeleton (shell and spines) of the sea urchin Echinometra Mathaei on human liver cancer cell line (HepG2). E. mathaei was collected from the intertidal zone of the Red Sea, Al-Ain Al-Sokhna, Egypt. HepG2 cells were treated with the MeOH extract for 24 and 48 hours to assess the cytotoxic activity and the potential molecular mechanisms. The expression levels of apoptosis-, autophagy-, and cell proliferation-related genes and/or proteins were measured. In addition, metabolomic profiling of the extract was performed by gas chromatography-mass spectrometry (GC-MS) analysis. The extract displayed cytotoxic activity on the liver cancer cells (IC50 29.47 and 14.97 µg/ml at 24 and 48 hours, respectively). It induced apoptosis, autophagy and inhibited cell proliferation of HepG2 cells. The metabolomic profiling showed that the extract is rich in fatty acids. These results demonstrated that the exoskeleton MeOH extract of E. mathaei induced apoptosis in HepG2 cells, confirming its cytotoxic effect. Therefore, a thorough investigation of its molecular target and structure is necessary to develop novel anticancer therapeutics from marine Sea urchins.

Rutaecarpine Induces Apoptosis via a Mitochondrial Membrane-Mediated Pathway in the Human Liver Cancer Cells (HepG2)

Rutaecarpine Induces Apoptosis via a Mitochondrial Membrane-Mediated Pathway in the Human Liver Cancer Cells (HepG2)

The Prognostic Significance of CTSV Expression in Patients with Hepatocellular Carcinoma.

Cathepsin V (CTSV) is a cysteine protease peptidase, which is typically upregulated in cancer and is associated with various oncogenic processes, such as angiogenesis, proliferation, apoptosis, and invasion. The study explored the role of CTSV in hepatocellular carcinoma (HCC) and its potential as a potential biomarker. This study collected tumor and peritumoral archived specimens from 180HCC patients who underwent surgical resection at Zhongshan Hospital, Fudan University (Shanghai, China) between 2009 and 2010. We extracted data from the TCGA and GEO databases and conducted differential expression analysis, univariate Cox regression, and Kaplan-Meier analysis. Ultimately, we determined that CTSV may emerge as a potential biomarker. Then, immunohistochemical staining for CTSV was performed on tumors and adjacent tissues of HCC patients, and a Cox proportional hazards model was constructed to evaluate the prognostic significance of CTSV expression levels. Applied functional enrichment analysis to reveal the underlying molecular mechanisms. Utilized ssGSEA enrichment analysis and TIMER2.0 algorithm to explore the correlation between CTSV expression and immune cells in HCC. In vitro and in vivo experiments were conducted using human liver cancer cell lines to further validate the clinical application value of CTSV. In this study, we observed that CTSV expression was notably elevated in HCC (P < 0.001), and identified a significant association between elevated CTSV expression and reduced overall survival rates in patients. In vitro and in vivo experiments indicated that CTSV knockdown could significantly inhibit the proliferation, migration, and invasion of liver cancer cells, and it was found that the combination of CTSV knockdown with PD-1 inhibitors might enhance the therapeutic effect of PD-1 inhibitors in HCC. CTSV serves as a standalone negative prognostic indicator and possesses clinical significance in HCC.

Determination of the interactions of a schiff base with different targets via molecular docking and cytotoxic activity studies

The pyrimidine nucleus is a vital pharmacophore that exhibits excellent pharmacological activity. A Schiff base containing a pyrimidine nucleus was obtained in two steps. This compound was assessed on the human prostate (PC3) and liver (HepG2) cancer cell lines using the MTT method. The prepared compound 3 was determined to have a high cytotoxic effect towards prostate cancer with an IC50 value of 9.32 μM. Since experimental prostate and liver cancer studies were examined, the molecular docking study's target structures were determined accordingly. In molecular docking studies, compound 3 interacted in silico with the crystal structure of the human HER2 kinase domain (PDB Id: 3PP0), the crystal structure of VEGFR-2 (PDB Id: 4ASD), and the crystal structure of EGFR tyrosine kinase (PDB Id: 4HJO), respectively. As a result of these interactions, binding energy values were calculated, and binding modes were determined. Additional in vitro and in vivo experiments targeting other cancer cell lines will provide deeper insight into the anticancer spectrum of this compound and new studies on this compound will be planned in the following years. Furthermore, optimizing and studying such pyrimidine-based Schiff base compounds for improved selectivity and potency against prostate cancer is also considered to be valuable as it may increase the potential to capture potential drug candidates.

Evaluating ECM stiffness and liver cancer radiation response via shear-wave elasticity in 3D culture models

BackgroundThe stiffness of the tumor microenvironment (TME) directly influences cellular behaviors. Radiotherapy (RT) is a common treatment for solid tumors, but the TME can impact its efficacy. In the case of liver cancer, clinical observations have shown that tumors within a cirrhotic, stiffer background respond less to RT, suggesting that the extracellular matrix (ECM) stiffness plays a critical role in the development of radioresistance.MethodsThis study explored the effects of ECM stiffness and the inhibition of lysyl oxidase (LOX) isoenzymes on the radiation response of liver cancer in a millimeter-sized three-dimensional (3D) culture. We constructed a cube-shaped ECM-based millimeter-sized hydrogel containing Huh7 human liver cancer cells. By modulating the collagen concentration, we produced two groups of samples with different ECM stiffnesses to mimic the clinical scenarios of normal and cirrhotic livers. We used a single-transducer system for shear-wave-based elasticity measurement, to derive Young’s modulus of the 3D cell culture to investigate how the ECM stiffness affects radiosensitivity. This is the first demonstration of a workflow for assessing radiation-induced response in a millimeter-sized 3D culture.ResultsIncreased ECM stiffness was associated with a decreased radiation response. Moreover, sonoporation-assisted LOX inhibition with BAPN (β-aminopropionitrile monofumarate) significantly decreased the initial ECM stiffness and increased RT-induced cell death. Inhibition of LOX was particularly effective in reducing ECM stiffness in stiffer matrices. Combining LOX inhibition with RT markedly increased radiation-induced DNA damage in cirrhotic liver cancer cells, enhancing their response to radiation. Furthermore, LOX inhibition can be combined with sonoporation to overcome stiffness-related radioresistance, potentially leading to better treatment outcomes for patients with liver cancer.ConclusionsThe findings underscore the significant influence of ECM stiffness on liver cancer’s response to radiation. Sonoporation-aided LOX inhibition emerges as a promising strategy to mitigate stiffness-related resistance, offering potential improvements in liver cancer treatment outcomes.

Morusin, a novel inhibitor of ACLY, induces mitochondrial apoptosis in hepatocellular carcinoma cells through ROS-mediated mitophagy

ObjectiveMorusin (Mor), a prenylated flavonoid isolated from the root bark of Morus alba L., exhibits potent anti-tumour effects; however, the molecular target of Mor is still not entirely clear. This study aimed to elucidate the mechanism of Mor against hepatocellular carcinoma (HCC) and identify potential molecular targets. MethodsMitochondrial function was assessed by measuring the mitochondrial membrane potential, mitochondrial ultrastructure, oxygen consumption, and ATP levels. Mor-induced mitophagy was confirmed using western blotting, immunofluorescence, and fluorescent probes. Transcriptomics, flow cytometry, western blotting, qRT-PCR and biochemical assays were used to reveal the molecular mechanisms and targets of Mor against HCC. We further validated the interaction between Mor and the target proteins using molecular docking and biolayer interferometry (BLI). The inhibitory effect of Mor in vivo was evaluated using a Hep3B murine xenograft model. ResultsMor significantly reduced the ATP citrate lyase (ACLY) expression and inhibited ACLY activity in HCC cells. BLI analysis demonstrated a direct interaction between Mor and the ACLY active domain. Mor-induced ACLY inhibition led to ROS accumulation in HCC cells, which caused mitochondrial damage, triggered PINK1/Parkin-mediated mitophagy, and ultimately induced mitochondrial apoptosis. We further verified that ROS is crucial in the apoptotic action of Mor through experiments regarding an ROS scavenger. Mor also significantly inhibited tumour xenograft growth in vivo. In addition, analysis of human liver cancer clinical samples revealed elevated ACLY levels positively correlated with histologic grade. ConclusionCollectively, our findings highlight Mor as a potent bioactive inhibitor of ACLY and a promising candidate for HCC therapy.

The loss of hepatitis B virus receptor NTCP/SLC10A1 in human liver cancer cells is due to epigenetic silencing.

HBV is a hepatotropic virus that infects human hepatocytes expressing the viral receptor hNTCP. Without effective antiviral therapy, chronic HBV infection poses a high risk of liver cancer. However, most liver cancer cell lines, including HepG2 and Huh7, do not support HBV infection due to the absence of hNTCP expression, and the mechanism underlying this defect remains unclear. This study demonstrates a significant reduction of hNTCP in hepatocellular carcinoma samples and HepG2 cells compared to normal liver tissues and primary human hepatocytes. Despite identical hNTCP genetic sequences, epigenetic analyses revealed a loss of the activating histone modification H3K27ac near the hNTCP transcription start site in cancer cells. Treatment with histone deacetylase inhibitors restored H3K27ac levels, reactivated hNTCP expression, and rendered HepG2 cells susceptible to HBV infection. These findings highlight the role of epigenetic modulation in hNTCP dysregulation, offering insights into hepatocarcinogenesis and its implications for chronic HBV infection.

Investigation of Cytotoxic Effects of Tragopogon reticulatus Against HepG2 and MCF-7 Cell Lines and Antioxidant, Antimicrobial Effects Against Some Microorganisms

Plants have been used for various purposes throughout history. This subject has received increasing attention in the field of traditional medicine in recent years. Tragopogon reticulatus, as a forage grass plant in turkey, is a species that stands out with rich vitamin and mineral content. The aim of this study was to investigate the antimicrobial, antioxidant, and cytotoxic activities of T. reticulatus. Two separate extracts were prepared at different concentrations using hexane and methanol solvents obtained from the T. reticulatus plant, and cytotoxic activity was evaluated by the MTT Assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method on human liver cancer (HepG2) and human breast cancer (MCF7) cell lines. The antioxidant activity of the extracts obtained from methanol and hexane was determined by the DPPH (2,2-diphenyl 1-picrylhydrazyl) radical scavenging capacity method. Antimicrobial activity was evaluated by the disk diffusion method, and bacteria such as Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Bacillus megaterium, and Candida albicans fungus were used in the study. The hexane extract obtained from the T. reticulatus plant showed a high cytotoxic activity of 44% in the MCF7 cell line, while it showed a 51% dead cell count in the HepG2 cell line. When the antioxidant results were evaluated, it was determined that the DPPH radical scavenging effect of the hexane extract exhibited the most effective activity at 19%. When the antimicrobial results were evaluated, it was determined that the concentrations prepared with hexane exhibited the most effective activity with a zone diameter of 16 mm against B. megaterium. These findings suggest that T. reticulatus plant extracts can be considered as a potential source. These results may shed light on future research into the biological effects of the plant and contribute to the development of new treatment methods.

The identification of potent dual-target monopolar spindle 1 (MPS1) and histone deacetylase 8 (HDAC8) inhibitors through pharmacophore modeling, molecular docking, molecular dynamics simulations, and biological evaluation.

Overexpression of monopolar spindle 1 (MPS1) and histone deacetylase 8 (HDAC8) is associated with the proliferation of liver cancer cells, so simultaneous inhibition of both MPS1 and HDAC8 could offer a promising therapeutic approach for the treatment of liver cancer. Dual-targeted MPS1/HDAC8 inhibitors have not been reported. A combined approach of pharmacophore modeling and molecular docking was used to identify potent dual-target inhibitors of MPS1 and HDAC8. Enzyme inhibition assays were performed to evaluate the optimal compound with the strongest inhibitory activity against MPS1 and HDAC8. The selectivity of MPH-5 for MPS1 and HDAC8 was assessed on a panel of 68 kinases and other histone deacetylases. Subsequently, molecular dynamics (MD) simulation verified the binding stability of the optimal compound to MPS1 and HDAC8. Ultimately, in vitro cellular assays and in vivo antitumor assays evaluated the antitumor efficacy of the most promising compound for the treatment of hepatocellular carcinoma. Six dual-target compounds (MPHs 1-6) of both MPS1 and HDAC8 were identified from the database using a combined virtual screening protocol. Notably, MPH-5 showed nanomolar inhibitory effect on both MPS1 (IC50 = 4.52 ± 0.21nM) and HDAC8 (IC50 = 6.07 ± 0.37nM). MD simulation indicated that MPH-5 stably binds to both MPS1 and HDAC8. Importantly, cellular assays revealed that MPH-5 exhibited significant antiproliferative activity against human liver cancer cells, especially HepG2 cells. Moreover, MPH-5 exhibited low toxicity and high efficacy against tumor cells, and it overcomes drug resistance to some extent. In addition, MPH-5 may exert its antitumor effects by downregulating MPS1-driven phosphorylation of histone H3 and upregulating HDAC8-mediated K62 acetylation of PKM2. Furthermore, MPH-5 showed potent inhibition of HepG2 xenograft tumor growth in mice with no apparent toxicity and presented favorable pharmacokinetics. The study suggests that MPH-5 is a potent, selective, high-efficacy, and low-toxicity antitumor candidate for the treatment of hepatocellular carcinoma.

Oncogenic plasmid DNA and liver injury agent dictates liver cancer development in a mouse model.

Primary liver cancer is an increasing problem worldwide and is associated with significant mortality. A popular method of modeling liver cancer in mice is plasmid hydrodynamic tail vein injection (HTVI). However, plasmid-HTVI models rarely recapitulate the chronic liver injury which precedes the development of most human liver cancer. We sought to investigate how liver injury using thioacetamide contributes to the pathogenesis and progression of liver cancer in two oncogenic plasmid-HTVI-induced mouse liver cancer models. Fourteen-week-old male mice received double-oncogene plasmid-HTVI (SB/AKT/c-Met and SB/AKT/NRas) and then twice-weekly intraperitoneal injections of thioacetamide for 6 weeks. Liver tissue was examined for histopathological changes, including fibrosis and steatosis. Further characterization of fibrosis and inflammation was performed with immunostaining and real-time quantitative PCR. RNA sequencing with pathway analysis was used to explore novel pathways altered in the cancer models. Hepatocellular and cholangiocellular tumors were observed in mice injected with double-oncogene plasmid-HTVI models (SB/AKT/c-Met and SB/AKT/NRas). Thioacetamide induced mild fibrosis and increased alpha smooth muscle actin-expressing cells. However, the combination of plasmids and thioacetamide did not significantly increase tumor size, but increased multiplicity of small neoplastic lesions. Cancer and/or liver injury up-regulated profibrotic and proinflammatory genes while metabolic pathway genes were mostly down-regulated. We conclude that the liver injury microenvironment can interact with liver cancer and alter its presentation. However, the effects on cancer development vary depending on the genetic drivers with differing active oncogenic pathways. Therefore, the choice of plasmid-HTVI model and injury agent may influence the extent to which injury promotes liver cancer development.