Human Liver Cancer Cells HepG2 Research Articles

Farnesiferol C Exerts Antiproliferative Effects on Hepatocellular Carcinoma HepG2 Cells by Instigating ROS-Dependent Apoptotic Pathway.

Farnesiferol C (Far-C) is a coumarin commonly extracted from Ferula asafetida and is popularly used as a traditional source of natural remedy. Liver cancer or hepatocellular carcinoma (HCC) has emerged as a major cause behind cancer burden, and limited therapeutic interventions have further aggravated the clinical management of HCC. In the present study, the authors tested the hypothesis that Far-C-instigated oxidative stress resulted in anti-proliferation and apoptosis instigation within human liver cancer HepG2 cells. The observations reported herewith indicated that Far-C exerted considerable cytotoxic effects on HepG2 cells by reducing the cell viability (p < 0.001) in a dose-dependent manner. Far-C exposure also resulted in enhanced ROS production (p < 0.01) which subsequently led to loss of mitochondrial membrane potential. Far-C-instigated oxidative stress also led to enhanced nuclear fragmentation and condensation as revealed through Hoechst-33342. These molecular changes post-Far-C exposure also incited apoptotic cell death which concomitantly led to significant activation of caspase-3 (p < 0.001). Furthermore, Far-C exhibited its competence in altering the expression of genes involved in apoptosis regulation (Bax, Bad, and Bcl2) along with genes exerting regulatory effects on cell cycle (cyclinD1) and its progression (p21Cip1 and CDK4). The evidence thus clearly shows the preclinical efficacy of Far-C against HepG2 cells. However, further mechanistic investigations deciphering the alteration of different pathways post-Far-C exposure will be highly beneficial.

Green Synthesis, Characterization, Antimicrobial and Anticancer Screening of New Metal Complexes Incorporating Schiff Base.

A Schiff base ligand of o-vanillin and 4-aminoazobenzene and its transition metal complexes of Ni(II), Co(II), Zn(II), Cu(II), Mn(II), and Zr(IV) were prepared under microwave irradiation as a green approach compared to the conventional method. The structures of new compounds have been characterized and elucidated via elemental and spectroscopic analyses. In addition, magnetic susceptibility, electron spin resonance, and electronic spectra of the synthesized complexes explained their geometrical structures. The thermal stability of Cu(II), Zn(II), and Zr(IV) complexes was studied by thermo-gravimetric analyses (TGA). Coats–Redfern and Horowitz–Metzger equations were used to calculate the thermal and dehydration decomposition activities of proposed structures kinetically. Surface morphologies of the solid compounds were imaged by scanning electron microscopy (SEM). The particle size of prepared complexes was measured by using a particle size analyzer at a diffraction angle of 10.9°. The geometry structures of the synthesized compounds were verified utilizing electronic spectra, ESR spectrum, and magnetic moment value. The newly synthesized compounds were screened for antimicrobial activity. Also, the anticancer activity of the free Schiff base ligand and its metal complexes were studied against two cell lines: human colon (HCT-116) and human liver cancer cells (HepG-2). The obtained results showed that the Cu(II) complex displayed the highest cytotoxic activity (IC50 = 18 and 22 μg/mL for HepG-2 and HCT, respectively) compared to the free Schiff base ligand.

Antitumor immunity and therapeutic properties of marine seaweeds-derived extracts in the treatment of cancer

Marine seaweeds are important sources of drugs with several pharmacological characteristics. The present study aims to evaluate the antitumor and antitumor immunological potentials of the extracts from the brown alga Padinapavonica and the red alga Janiarubens, inhibiting the Egyptian marine coasts. Hep-G2 cell lines were used for assessment of the antitumor efficacy of Padinapavonica and Janiarubens extracts in vitro, while Ehrlich ascites carcinoma (EAC) cells were applied to gain more antitumor immunity and antitumor insights of P.pavonica and J.rubens extracts in vivo. In vitro antitumor potentials of P.pavonica and J.rubens extracts were analyzed against human liver cancer Hep-G2 cells by MTT and trypan blue exclusion assays. In vivo antitumor immunological potentials of P.pavonica and J.rubens extracts at low, high, and prophylactic doses were analyzed by blood counting and flow cytometry in mice challenged with Ehrlich ascites carcinoma (EAC) cells. In vitro results revealed that P.pavonica and J.rubens extracts caused significant decreases in the number and viability of Hep-G2 cells in a dose-dependent manner as compared to untreated Hep-G2 cells or Cisplatin®-treated Hep-G2 cells. In vivo findings showed that P.pavonica and J.rubens extracts at low, high, and prophylactic doses significantly reduced the number and viability of EAC tumor cells accompanied by increases in EAC apoptosis compared to naïve EAC mouse. Additionally, P.pavonica and J.rubens extracts at low and prophylactic doses remarkably increased both the total WBC count and the relative numbers of lymphocytes and decreased the relative numbers of neutrophils and monocytes. Flow cytometric analysis showed that P.pavonica and J.rubens extracts at the treatment and the prophylactic doses resulted in a significant increase in the phenotypic expressions of CD4+ T, CD8+ T, and CD335 cells compared to naïve EAC mouse. Overall, both extracts P.pavonica and J.rubens possess potential antitumor and antitumor immunological effects with less toxicity, opening new approaches for further studies of the chemical and biological mechanisms behind these effects.

Antioxidant and chemoprotective peptides from simulated gastrointestinal digested (SGID) protein hydrolysate of Pyropia yezoensis against acetaminophen-induced HepG2 cells.

Excessive production of reactive oxygen and nitrogen species may result in oxidative damage to tissues and organs. Oxidative stress is a pathological mechanism that contributes to the initiation and progression of liver injury. In the present study, antioxidative peptides purified from simulated gastrointestinal-digested (SGID) protein hydrolysate of Pyropia yezoensis, showed significant antioxidant activity and also showed a protective effect against acetaminophen (N-acetyl-p-aminophenol, APAP) -induced injury in HepG2 (human liver cancer cells) cells. The antioxidant activity was increased in a dose-dependent manner. Higher cell viability (73.26 ± 0.9%) and decreasing NO levels (107.6 ± 8.9%) were observed in 15 mM APAP-induced cells when treated with the concentration of (100 μg ml−1) Pyropia peptide. Py. (pep). The sequences of the eight identified peptides present in the active fractions of the protein hydrolysate included hydrophobic and aromatic amino acids, which may have been responsible for their chemoprotective and antioxidant activities. Results indicated that the treatment with the Pyropia—peptides significantly promoted the proliferation of HepG2 cells, protecting them against APAP-mediated injury, and showed a significant antioxidant capacity. This study revealed that the Py. (pep) will be beneficial in treating drug-induced oxidative stress and liver damage conditions. Py. (pep) can also serve as a better alternative for synthetic antioxidant drugs.

GCMS ANALYSIS ON ANDROGRAPHIS PANICULATA SEED EXTRACT AND ITS ANTICANCER ACTIVITY

Objective: Andrographis paniculata was an ancient medicinal herb recorded in Ayurvedic system of medicine from long times. The present study is to determine the bioactive phytocomponents present in Andrographis paniculata seed, extraction done using ethanol and Human Liver Cancer (HepG2) cells are used to prove the anticancer activity. Methods: Gas Chromatography-Mass Spectroscopy (GCMS) analysis is used for determining the bioactive constituents of the Andrographis paniculata seed. To prove the in vitro anticancer activity, the MTT assay was done on Human Liver Cancer (HepG2) cells. Results: GCMS analysis of Andrographis paniculata seed extract result determined the presence of 15 active bio components. These various potent bioactive compounds have been proved to possess various pharmacological activities, viz., antimicrobial, antidiabetic, antioxidant, anti-inflammatory and anticancer properties, which were supported by previous findings. The natural compounds with therapeutic effects on traditional medicine have paved the way for evaluating the Andrographis paniculata seeds for cytotoxic activity on HepG2 cell lines. The ethanol extract of Andrographis paniculata seeds was used on HepG2 to evaluate the cytotoxicity; Cisplatin (15 µg/ml) is used as standard with various concentration like 200, 100, 50 and 25 µg/ml were used. The bioactive compounds present in Andrographis paniculata seed extract revealed significant cytotoxicity activity against HepG2 cells with IC50 value of 103.03µg/ml. Conclusion: The cytotoxic effect was found to be concentration-dependent and increased concentration of Andrographis paniculata seed extract showed increased cytotoxicity. The result suggest Andrographis paniculata seed extract have bioactive agents which when treated on hepatocellular carcinoma cells poses excellent anticancer activity.

Anticancer activity of G4-targeting phenoxazine derivatives in vitro

G4-stabilizing ligands are now being considered as anticancer, antiviral and antibacterial agents. Phenoxazine is a promising scaffold for the development of G4 ligands. Here, we profiled two known phenoxazine-based nucleoside analogs and five new nucleoside and non-nucleoside derivatives against G4 targets from telomere repeats and the KIT promoter region. Leading new derivatives exhibited remarkably high G4-stabilizing effects (comparable or superior to the effects of the commonly used selective G4 ligands PDS and NMM) and selectivity toward G4s over duplex (superior to BRACO-19). All phenoxazine-based ligands inhibited cellular metabolic activity. The phenoxazine derivatives were particularly toxic for lung adenocarcinoma cells A549' and human liver cancer cells HepG2 (CC50 of the nucleoside analogues in the nanomolar range), but also affected breast cancer cells MCF7, as well as immortalized fibroblasts VA13 and embryonic kidney cells HEK293t (CC50 in the micromolar range). Importantly, the CC50 values varied mostly in accordance with G4-binding affinities and G4-stabilizing effects, and the phenoxazine derivatives localized in the cell nuclei, which corroborates G4-mediated mechanisms of action.

Novel specific cardiac homing peptides to target failing cardiomyocytes

Heart failure (HF) is one of the most prevalent cardiovascular diseases worldwide with high mortality. Although β-blockers are now the cornerstone of HF treatment, they are ineffective in patients’ subpopulations and may cause several side effects. Thus, the development of novel drug delivery systems directly targeting the myocardium in a tissue-specific manner is a promising approach. Two ligands have been characterized to transduce cardiomyocytes (CMs) in vitro as well as cardiac tissue in vivo: the cardiac targeting peptide (CTP) and the primary cardio myocyte (PCM) peptide. However, very little is known about their uptake mechanisms, targeting efficacies, and intracellular distribution. Characterization and optimization of the CTP and PCM peptides through a structure-activity relationship study. Over 100 variants of CTP and PCM peptides have been engineered and screened. The targeting capacity of peptides was evaluated using flow cytometry on neonatal rat ventricular CMs (NRVC) cultured with 6-carboxyfluorescein conjugated peptides. Other cells lines, including human lung adenocarcinoma (A549) and human liver cancer (HepG2) cells, were used to assess the cardiospecificity of the new peptides. Peptide uptake was also monitored by confocal microscopy imaging in NRVC and adult rat ventricular CMs (ARVC), as well as in the ex vivo model. We have generated specific cardiac homing peptides-1 and 2 (i.e., SCHoP-1 and SCHoP-2) with improved properties and increased ability to target and transduce CMs specifically, compared to the parent CTP and PCM peptides. Flow cytometry assays support that SCHoP-1 and SCHoP-2 significantly increase cardiac targeting in living NRVC by 4-fold over the CTP and PCM peptides. No cellular toxicity was observed. SCHoP-1 and SCHoP-2 target preferentially CMs, over A549 and HepG2 cells. We confirmed that they internalize in NRVC and in ARVC localized in the cytosol. CMs internalization is also increased in ex vivo hearts. SCHoP-1 and SCHoP-2 open the way to explore their potential as a targeting moiety to address a therapeutic agent specifically to failing CMs for the treatment of HF.

Solid Lipid Nanoparticles of Lepidium Sativum L Seed Extract: Formulation, Optimization and In vitro Cytotoxicity Studies.

The current study focused on important bioactive compounds in plants that make them pharmacologically valuable. Therefore, this study was aimed to develop Lepidium sativum (L. sativum) seed extract loaded solid lipid nanoparticles and explore its cytotoxic effect on human liver cancer cells (HepG2 cells). The ethanolic extract of L. sativam used to develop L. sativum seed extract loaded solid lipid nanoparticles (SLNs) was analyzed by gas chromatography-mass spectrometry, thin-layer chromatography (TLC) and high-performance thin-layer chromatography (HPTLC) for phytochemical profiling. The L. sativum seed extract loaded SLNs were efficaciously prepared by the nanoprecipitation method and screened on the basis of physicochemical properties. The L. sativum seed extract loaded SLN-2 was characterized using various parameters like particle size (237.1±0.104), % entrapment efficiency (80±1.15), zeta potential (42.1±0.102) and % drug release (45% at the end 8 hours and release the entire amount in 12 h). The SLN-2 formulation was optimized based on the recipient factor, and SLN-2 was used to further evaluate the in vitro cytotoxicity of HepG2 cells in a dose-dependent manner by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. The IC50 value of SLN2 was 52.37 ug/ml and sub IC50 26.1 ug/ml at 24 h and 48 h, respectively. Thus, we concluded that L. sativum extract loaded SLN-2 could act as an alternative therapy, possibly controlling therapeutic action by making a substantial reduction in side effects.

High-resolution 3D printing of xanthan gum/nanocellulose bio-inks

The current study provides a comprehensive rheology study and a survey on direct ink writing of xanthan gum/cellulose nanocrystal (XG/CNC) bio-inks for developing 3D geometries that mimic soft tissue engineering scaffolds' physical and mechanical properties. The presence of CNC was found to be a critical prerequisite for the printability of XG bio-inks; accordingly, the hybrid XG/CNC bio-inks revealed the excellent viscoelastic properties that enabled precise control of hydrogel shaping and printing of lattice structures composed of up to eleven layers with high fidelity and fair resolution without any deformation after printing. The lyophilized 3D scaffolds presented a porous structure with open and interconnected pores and a porosity higher than 70%, vital features for tissue engineering scaffolds. Moreover, they showed a relatively high swelling of approximately 11 g/g, facilitating oxygen and nutrient exchange. Furthermore, the elastic and compressive moduli of the scaffolds that enhanced significantly upon increasing CNC content were in the range of a few kPa, similar to soft tissues. Finally, no significant cell cytotoxicity was observed against human liver cancer cells (HepG2), highlighting the potential of these developed 3D printed scaffolds for soft tissue engineering applications.

Probing the Mechanism of Action of Cry41Aa on HepG2 through the Establishment of a Resistant Subline.

Cry41Aa, also called parasporin-3, belongs to a group of toxins from the entomopathogenic bacterium Bacillus thuringiensis that show activity against human cancer cells. Cry41Aa exhibits preferential cytocidal activity towards HL-60 (human promyelocytic leukaemia cells) and HepG2 (human liver cancer cells) cell lines after being proteolytically activated. To better understand the mechanism of action of Cry41Aa, we evolved resistance in HepG2 cells through repeated exposure to increasing doses of the toxin. Concentrations of Cry41Aa that killed over 50% of the parental HepG2 cells had no significant effect on the viability of the resistant cells and did not induce either pore formation or p38 phosphorylation (both characteristic features of pore-forming toxins). Preliminary RNA sequencing data identified AQP9 as a potential mediator of resistance, but extensive investigations failed to show a causal link and did not support an enhanced cell repair process as the resistance mechanism.

Exploring the Therapeutic Potentials of Exopolysaccharides Derived From Lactic Acid Bacteria and Bifidobacteria: Antioxidant, Antitumor, and Periodontal Regeneration.

The metabolites of lactic acid bacteria (LAB) and bifidobacteria (Bb) have recently received a lot of attention due to their ability to protect interactions in blood and tissues, as well as their biodegradability and biocompatibility in human tissue. Exopolysaccharides (EPS) derived from bacteria have a long history of use in therapeutic and other industrial applications with no adverse effects. In this regard, EPSs were isolated and characterized from LAB and Bb culture supernatants to determine their antioxidant, antitumor, and periodontal regeneration properties. The antioxidant capacity of the EPSs varied with concentration (0.625–20 mg/ml). The highest antioxidant activity was found in LAB: Streptococcus thermophiles DSM 24731-EPS1, Lactobacillus delbrueckii ssp. bulgaricus DSM 20081T-EPS5, Limosilactobacillus fermentum DSM 20049-EPS6, and Bb; Bifidobacterium longum ssp. longum DSM 200707-EPS10. Human breast cancer cells (MCF7), human colon cancer cells (CaCo2), human liver cancer cells (HepG2), and human embryonic kidney 293 (HEK 293) cells were used as controls to assess the antitumor properties of the selected EPSs. According to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) assay, EPS5 had the highest cytotoxicity against MCF7, CaCo2, and HepG2, with IC50 values of 7.91, 10.69, and 9.12 mg/ml, respectively. Lactate dehydrogenase (LDH) activity was significantly higher in cell lines treated with EPS5-IC50 values compared to other EPSs-IC50 values (p < 0.05). Real time (RT)-PCR results showed that EPS5 treatment increased Bax, Caspase 8, Caspase 3, and p53 gene expression. The expression of the BCL2, MCL1, and Vimentin genes, on the other hand, was reduced. The MTT test was used to examine the effect of EPS5 on the viability of human periodontal ligament fibroblast cells (hPDLFCs), and it was discovered that EPS5 increased hPDLFC viability. According to high-performance liquid chromatography (HPLC) analysis, galactose made up 12.5% of EPS5. The findings of this study pave the way for the use of EPS, which hold great promise for a variety of therapeutic purposes such as antioxidant, antitumor, and periodontal regeneration.

Prediction of the potential mechanism of compound gingerol against liver cancer based on network pharmacology and experimental verification.

To explore gingerol's potential mechanism for treating liver cancer using network pharmacology and molecular docking technology and to conduct in-vitro experiments of human liver cancer cell HepG2 to verify important signalling pathways. We obtained potential targets of gingerol derivatives (6-gingerol, 8-gingerol and 10-gingerol) from PubChem and SwissTargetPrediction websites and collected related targets for liver cancer with the help of GeneCards. We performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis on key targets using the DAVID data platform and combined with Cytoscape 3.7.1 software to construct a component-target-signal pathway interaction map to study its mechanism of action. Subsequently, the components and key proteins were molecularly docked through Autodock Vina software. Finally, the important signal pathways were verified by HepG2 cell in-vitro experiments. A total of 318 drug targets were screened for gingerol derivatives, and 2509 gene targets related to liver cancer were collected. The Venn diagram showed that there were 104 intersection targets between gingerol derivatives and liver cancer. Module analysis results show that these intersection targets can be divided into 5 modules and 49 nodes. Bioinformatics analysis found that GO obtained 20 important functional items including cancer cell proliferation, protein kinase activity, phosphotransferase activity and kinase activity; KEGG enrichment analysis yielded a total of 20 key signal pathways including the PI3K-Akt signalling pathway. The results of molecular docking show that the binding energy of gingerol derivatives has good binding activity with PI3K and Akt. In-vitro experimental results show that gingerol derivatives and compound gingerol (compound gingerol is composed of 6-gingerol, 8-gingerol and 10-gingerol in a ratio of 7:1.5:1.5) can produce HepG2 cell proliferation inhibition, and each administration group can significantly increase the apoptosis rate of HepG2 cells and the fluorescence intensity of the nucleus and block the cell cycle in the S phase; the results of Western Blot and real-time quantitative PCR show that gingerol derivatives and compound gingerol can down-regulate the expression of Akt and p-Akt and up-regulate the expression of Bax/Bcl-2. And the effect of compound gingerol is more obvious than that of gingerol derivatives. The results of network pharmacology and experimental validation suggest that gingerol derivatives and compound gingerol can act against liver cancer by acting on the PI3K-Akt signalling pathway.

Induction of Bax and Activation of Caspases Hydro Ethanoic Leaf Extract of Citrullus colocynthis(L)- Mediated Apoptosis in Human Liver Cancer Cells Hep G2

Introduction: Citrullus colocynthis is a desert plant of the cucurbitaceae family. C. colocynthis grows in many countries with semi dry lands and that seeds are rich in oil which can encourage their sustainable agriculture to produce an economic biodiesel with a competitive price. Apoptosis, also termed as programmed cell death this is a key regulator of tissue homeostasis.&#x0D; Aim of the Study: To study the induction of bax and activation of caspases hydro ethanolic leaf extract of Citrullus colocynthis (L)- mediated apoptosis in human liver cancer cells Hep G2.&#x0D; Materials and Methods: In the present study, a human liver cancer cell line was procured from the National Centre for Cell science (NCCS), Pune, India. The cytotoxic potential of plant extract was analysed by MTG assay. mRNA expression of BaxmRNA and caspase-mRNA were analysed by Real-Time PCR using gene specific primers and a comparative CT method was used to analyse expression of genes. The data were analysed statistically using Graph pad prism software Version 5.&#x0D; Results: MTT analysis showed that Citrullus colocynthis a significant increase in proliferation of human liver cancer cells (P&lt;0.05) compared to untreated cells. However, 400 and 500 /g of the extract showed maximum effect in controlling cell growth. Results of the &lt;mRNA expression analysis showed that Citrullus colocynthis reduced the expression of BaxmRNA/ caspase-mRNA in human liver cancer cells at 400 and 500 /g (P&lt;0.05).&#x0D; Discussion: The anticancer effect of Citrullus colocynthis has been via a variety of pathways which includes apoptotic pathway, anti-inflammatory traits, and antioxidant pathway, etc [1]. Effect of HEAM on the mRNA level of tumor related genes (p53, Bcl-2), inflammatory (IL-1β, IL-6 and c-jun) and anti-inflammatory (IL-4) cytokines. Representative photographs of ethidium-bromide-stained 2% agarose gels for each gene after 7 months of treatment.&#x0D; Conclusion: The hydroethanolic leaf extract obtained from the Citrullus colocynthis exhibited anti-proliferative effect on human liver cancer cells HepG2 cell line by affecting the expression of the Bax mRNA and Bax mRNA and hindering the cell viability. Hence, it can be a great scope for using the extract into medicinal properties with further studies to improve herbal medicine for cancer over expensive drugs.

Synthesis, anti‐proliferative activities, docking studies, and computational calculations of novel isonicotinic mixed complexes

AbstractIn this investigation, we studied the mixing of two ligands, isonicotinohydrazide (L1) and 3‐hydroxyisonicotinic acid (L2), with different metal ions to afford the corresponding stable metal complexes. Consequently, intensive physico‐chemical analysis including elemental analysis, UV–visible, Fourier transform infrared (FT‐IR), thermogravimetric analysis (TGA), mass, 1H nuclear magnetic resonance (NMR), X‐ray diffraction (XRD), magnetic, and molar conductance measurements has been performed. The thermal stability of all complexes was determined using TGA, and the kinetic parameters were obtained using the Coats–Redfern method. Both the degree of crystallinity and the unit cell of all complexes were determined utilizing XRD studies. Furthermore, these metal complexes exhibited excellent anti‐proliferative activity against human breast cancer (MCF‐7) and human liver cancer (HepG2) cell lines against the doxorubicin drug. The Co(II) complex outperformed all other metal complexes in terms of activity, which was confirmed by molecular docking stimulation via the binding energy with amino acids in a structural complex of Kaposi's sarcoma‐associated herpesvirus (KSHV) thymidylate synthase (PDBID:5H38) and the ternary complex of KRIT1 bound to both the Rap1 GTPase and the Heart of Glass (HEG1) cytoplasmic tail (PDBID:4hdq) and shortage. Moreover, these metal complexes were optimized via theoretical investigation through a density functional theory (DFT)/B3LYP/LANL2DZ basis set to confirm the stability of these metals theoretically and detect frontier molecular orbitals (FMO), electrostatic potential (ESP), energy gap, and physical computational calculations. Eventually, the electrochemical features were explored using cyclic voltammetry (CV) and electrochemical impedance spectra (EIS).

MiR-205 Reverses MDR-1 Mediated Doxorubicin Resistance via PTEN in Human Liver Cancer HepG2 Cells.

Objective The aim of the recent study was to investigate the effects of miR-205 on reversing Doxorubicin (DOX) resistance, as chemotherapeutic agents through up-regulation of PTEN in human liver cancer HepG2 cells. Materials and Methods In this experimental study, the drug resistance in liver cancer cells via drug efflux inhibition and enhancing apoptosis by the regulation of PTEN and multi-drug resistance/ P-glycoprotein (MDR/P-gp) expression was revealed. Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, effect of DOX on cell proliferation was evaluated after miR-205 transfection in HepG2 and HepG2/DOX cells. Activity of P-gp on drug efflux was measured by the Rhodamine 123 (Rho-123) assay. PTEN mRNA expression levels were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and flow cytometry was used to measure the apoptotic ratio of HepG2/DOX cells. Results miR-205 overexpression considerably inhibited the HepG2/DOX cells viability (P<0.05). qRT-PCR results revealed that PTEN is a pivotal regulator in PI3K/Akt/P-gp axis. Overexpression miR-205 resulted in up-regulation PTEN and ultimately down-regulation of P-gp. This inhibits drug resistance, proliferation and induces apoptosis in HepG2/DOX cells (P<0.05). Whilst, treatment with 10 μM of special inhibitors, including LY294002 (PI3K) or PD098059 (MAPK), increased Rho 123-associated MFI, treatment with 10 μM of SF1670 (PTEN) almost abolished the effect of miR-205 overexpression (P<0.05). Finally, we found that miR-205 was down-regulated in HepG2/DOX cells, and its overexpression led to enhancing apoptosis with re-sensitization of HepG2/DOX cell lines to DOX through PTEN/PI3K/ Akt/MDR1 pathway. ConclusionThese findings may introduce miR-205 as a predictive biomarker and a potential treatment target for liver cancer therapy during MDR.

Hyper-branched multifunctional carbon nanotubes carrier for targeted liver cancer therapy

The systemic toxicity of anticancer drugs regularly restricts the use of conventional chemotherapy to treat cancer. In this study, the limitations overcome by profitably fabricating a multifunctional nanocarrier system to carry the anticancer drug into the specific location of the cancer cells. The polyethylene glycol (PEG) was functionalized in the carboxylated multiwalled carbon nanotubes (MWCNT-COOH) through an esterification reaction (MWCNT-PEG). The targeting ligand of folic acid (FA) was covalently bonded with hyperbranched poly-L-lysine (HBPLL) using adipic acid (AA) as a cross-linking agent. Doxorubicin (DOX), an anticancer drug, was effectively loaded on MWCNT-PEG-AA-HBPLL-FA carrier loading, and in-vitro drug release was investigated by UV–Vis spectrophotometer. The chemical functionalization, morphological properties, crystalline nature, surface charge, and thermal stability of the synthesized materials were studied by FT-IR, FE-SEM, HR-TEM, DLS, and TGA techniques. In-vitro cytotoxicity and anticancer properties of DOX-loaded nanocarrier were studied in human liver cancer (HepG2) cells and human embryonic kidney (HEK293) cells. The activities of caspases (caspase −3, −8 & −9) were analyzed using luminometry. The intrinsic apoptosis pathway proteins (Bcl-2 & BAX) were determined by western blot and RT-PCR analysis. The synthesized DOX-loaded nanocarriers exhibited increased cytotoxicity and apoptosis in liver HepG2 cells. The results suggest that the DOX-loaded nanocarrier possesses strong anticancer properties and could be an applicable and potential drug carrier for liver cancer chemotherapy.

Hexokinase 2 Inhibition and Biological Effects of BNBZ and Its Derivatives: The Influence of the Number and Arrangement of Hydroxyl Groups.

Hexokinase 2 (HK2), an enzyme of the sugar kinase family, plays a dual role in glucose metabolism and mediating cancer cell apoptosis, making it an attractive target for cancer therapy. While positive HK2 expression usually promotes cancer cells survival, silencing or inhibiting this enzyme has been found to improve the effectiveness of anti-cancer drugs and even result in cancer cell death. Previously, benitrobenrazide (BNBZ) was characterized as a potent HK2 inhibitor with good anti-cancer activity in mice, but the effect of its trihydroxy moiety (pyrogallol-like) on inhibitory activity and some cellular functions has not been fully understood. Therefore, the main goal of this study was to obtain the parent BNBZ (2a) and its three dihydroxy derivatives 2b–2d and to conduct additional physicochemical and biological investigations. The research hypothesis assumed that the HK2 inhibitory activity of the tested compounds depends on the number and location of hydroxyl groups in their chemical structure. Among many studies, the binding affinity to HK2 was determined and two human liver cancer cell lines, HepG2 and HUH7, were used and exposed to chemicals at various times: 24 h, 48 h and 72 h. The study showed that the modifications to the structures of the new BNBZ derivatives led to significant changes in their activities. It was also found that these compounds tend to aggregate and exhibit toxic effects. They were found to contribute to: (a) DNA damage, (b) increased ROS production, and (c) disruption of cell cycle progression. It was observed that, HepG2, occurred much more sensitive to the tested chemicals than the HUH7 cells; However, regardless of the used cell line it seems that the increase in the expression of HK2 in cancer cells compared to normal cells which have HK2 at a very low level, is a serious obstacle in anti-cancer therapy and efforts to find the effective inhibitors of this enzyme should be intensified.

First and efficient bio-nano composite, SnO2/Calcite based on Cypress leaves and eggshell wastes, for cytotoxic effects on HepG2 liver cancer cell lines and its antioxidant and antimicrobial activity

The present study aimed to explain the influence of novel SnO2/Calcite bio-nano composite on HepG2 cell line, and its antioxidant, and antimicrobial properties. In this study, green fabrication of SnO2/Calcite bio-nano composite was synthesized based on cypress leaves and eggshell wastes, characterized structurally by SEM, TEM, XRD, EDAX, elemental analysis, and FT-IR for more information about this composite. We study the cytotoxic potential of SnO2/Calcite bio-nano composite against human liver cancer HepG2 cell line. In vitro free radical scavenging, total antioxidant capacity, and antimicrobial activity of this bio-nano composite were investigated. SnO2/Calcite bio-nano composite was displayed anti-cancer, antioxidant, and antimicrobial properties successfully. Also, the results suggest that SnO2/Calcite bio-nano composite holds more significant potential than calcite and SnO2 nanoparticles against bacteria and fungi. These results displayed that SnO2/Calcite bio-nano composite might be used for the design of novel anti-proliferative and antimicrobial bioagents.

Microwave-Assisted Extraction of Anticancer Flavonoid, 2',4'-Dihydroxy-6'-methoxy-3',5'-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits.

2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethyl chalcone (DMC) is a biological flavonoid that is present in the fruits of Syzygium nervosum (Ma-Kiang in Thai). Microwave-assisted extraction (MAE), which utilizes microwave radiation to heat the extraction solvent quickly and effectively, was used to recover DMC-rich extract from Syzygium nervosum fruit. To determine the DMC content, a highly accurate and precise HPLC technique was developed. The influences of MAE conditions, including the solid–liquid ratio, microwave power, and microwave duration on the content of DMC, were sequentially employed by a single factor investigation and response surface methodology (RSM) exploratory design. The predicted quadratic models were fitted due to their highly significant (p < 0.0001) and excellent determination coefficient (R2 = 0.9944). The optimal conditions for producing DMC-rich extract were a ratio of sample to solvent of 1:35 g/mL, a microwave power of 350 W, and a microwave time of 38 min. Under the optimal MAE setting, the DMC content reached 1409 ± 24 µg/g dry sample, which was greater than that of the conventional heat reflux extraction (HRE) (1337 ± 37 µg/g dry sample) and maceration (1225 ± 81 µg/g dry sample). The DMC-rich extract obtained from MAE showed stronger anticancer activities against A549 (human lung cancer cells) and HepG2 (human liver cancer cells) than the individual DMC substance, which makes MAE an effective method for extracting essential phytochemicals from plants in the nature.

Synthesis of ultrasonic assisted co-precipitated Ag/ZnO nanorods and their profound anti-liver cancer and antibacterial properties

The current cancer Survey reports liver cancer is the second leading cause of cancer related death worldwide. Active liver cancer research is underway as several new therapies are emerging to treat it, with more showing promise. Our present study aims at preparation of silver/zinc oxide nanocomposites for potential applications in antibacterial, anti-inflammatory, anti-cancerous, against human liver cancer cells (hepG2). Silver ions added Zinc oxide (Ag/ZnO) have been synthesized by ultrasonic assisted co-precipitation method. It is found that 3% of silver ions added ZnO were precipitated as nanoplates. And interestingly the observation shows when the concentration of the silver ions increased to 5%, it reduced as nanorods. X-Ray diffraction, SAED patterns and Fourier Transform Infrared Spectrum confirms that silver ions are well attached in ZnO lattice. Ag/ZnO possesses good absorption in the UV region. The reduction in average particle size due to the increase of silver ion concentration in ZnO lattice was confirmed by dynamic light scattering (DLS).SEM and TEM studies witnessed the micro structural transformation from nanoplates (3% Ag) to the rod-like structure on (5% Ag).Ag/ZnO displayed strong antibacterial activity against different pathogens. The obtained results are comparable with standard antibiotics against pathogenic bacteria. Anti-inflammatory study reveals that Ag/ZnO prevents inflammation. Dose dependent response of Ag/ZnO against human liver cancer cells (HepG2) indicates its greater efficacy in killing cancer cells. The selective silver ion concentration (5%) in ZnO have reduced its cross-sectional area with respect to volume. This property helps the silver ion/ZnO nanorods to penetrate efficiently into the affected cells and fight against bacteria, suppress cell inflammation, eradicate the growth of liver cancer cells; which shows feasibility of this composites in further research for therapeutic applications.