Hepatocellular Liver Carcinoma Cell Line Research Articles

Lysine demethylase 5C epigenetically reduces transcription of ITIH1 that results in augmented progression of liver hepatocellular carcinoma.

Lysine demethylase 5C (KDM5C) is a member of the KDM family of demethylases and has been reported as a cancer driver. This study aimed to probe the function of KDM5C in the development of liver hepatocellular carcinoma (LIHC) and the molecules of action. According to data from publicly accessible bioinformatic databases, KDM5C is highly expressed in LIHC and associated with poor patient prognosis. High expression of KDM5C was detected in acquired LIHC cell lines. Downregulation of KDM5C weakened proliferation, migration, invasiveness, and resistance to death of the LIHC cells in vitro, and it reduced growth of the xenograft tumors in nude mice. Inter-alpha-trypsin inhibitor heavy chain 1 (ITIH1) was predicted as a downstream gene negatively regulated by KDM5C. KDM5C-regulated H3K4me1 modification at the promoter region of ITIH1, inducing its transcriptional inactivation. Further downregulation of ITIH1 in cancer cells blocked the functions of KDM5C silencing and restored the malignant behaviors of LIHC cells. The activity of the PI3K/AKT signaling was decreased following KDM5C downregulation but recovered upon ITIH1 silencing. In conclusion, this study suggested that KDM5C epigenetically reduces ITIH1 and activates the PI3K/AKT signaling pathway to promote LIHC progression.

Effect of rosiglitazone on 20-hydroxyeicosatetraenoic acid levels and CYP4F2 expression in HepG2 cells


 
 
 
 Purpose: To determine the effect of rosiglitazone on the levels of the cardiotoxic arachidonic acid metabolite, 20-hydroxyeicosatetraenoic acid (20-HETE), in the human liver hepatocellular carcinoma cell line, HepG2.
 Methods: HepG2 cells were treated with thiazolidinedione rosiglitazone and the mRNA and protein expressions of cytochrome P450 4F2 (CYP4F2) responsible for synthesizing 20-HETE were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. The levels of 20-HETE were evaluated using liquid chromatography/mass spectrometry (LC-MS).
 Results: Rosiglitazone significantly increased the levels of CYP4F2 mRNA and protein when compared with the control group (p < 0.05). This was correlated with significantly increased 20-HETE levels in the culture medium of rosiglitazone-treated cells in a dose-dependent manner (p < 0.05). The PPARγ antagonist, GW9662, significantly repressed the increased production of 20-HETE and CYP4F2 mRNA protein (p < 0.05).
 Conclusion: Rosiglitazone increases the synthesis of 20-HETE via activation of PPARγ receptor and upregulation of CYP4F2. These findings may provide an additional explanation, at least in part, for the unwanted side effects of rosiglitazone on the cardiovascular system.
 
 
 

NLRP3 promotes immune escape by regulating immune checkpoints: A pan-cancer analysis

NLRP3 plays a pathogenic role in tumorigenesis by regulating innate and acquired immunity, apoptosis, differentiation, and intestinal microbes in tumors. Our research aimed to investigate the role of NLRP3 in pan-cancers based on multi-omics data in the TCGA database. Most types of tumors showed increased expression of NLRP3. Among them, the overexpressed NLRP3 in liver hepatocellular carcinoma (LIHC) and ovarian cancer (OV) indicated worse overall survival (OS). Further analysis also confirmed overexpressed NLRP3 in colon cancer (COAD) indicated a high probability of microsatellite instability (MSI) and low tumor mutational burden (TMB), which indicated a better response to immune checkpoint inhibitors (ICIs). Interestingly, overexpression of NLRP3 was closely related to high infiltration of immune cells (T cells, B cells, etc.) and overexpressed immune checkpoints (PD-1, PD-L1, LAG3, etc.). These results demonstrated NLRP3 promoted immune escape in cancers. Finally, we investigated the expression of various immune checkpoints by treating NLRP3 inhibitor MCC950 during the co-culture of peripheral blood mononuclear cells (PBMC) and LIHC cell line Hep3B. MCC950 significantly repressed the expression of PD-L1 and LAG3, and promoted the apoptosis rate of Hep3B. In conclusion, our research demonstrated the role of NLRP3 in pan-cancer, especially in LIHC. Inhibition of NLRP3 promoted the killing effect of T cells to cancer cells by repressing the expression of immune checkpoints.

Identification of SHCBP1 as a potential biomarker involving diagnosis, prognosis, and tumor immune microenvironment across multiple cancers

Shc SH2-domain binding protein 1 (SHCBP1), a protein specific binding to SH2 domain of Src homolog and collagen homolog (Shc), takes part in the regulation of various signal transduction pathways, which has been reported to be associated with tumorigenesis and progression. However, the pathological mechanisms are not completely investigated. Thus, this study aimed to comprehensively elucidate the potential functions of SHCBP1 in multiple cancer types. The comprehensive analyses for SHCBP1 in various tumors, including gene expression, diagnosis, prognosis, immune-related features, genetic alteration, and function enrichment, were conducted based on multiple databases and analysis tools. SHCBP1 was upregulated in most types of cancers. The results of qRT-PCR had confirmed that SHCBP1 mRNA was significantly upregulated in lung adenocarcinoma (LUAD) and liver hepatocellular carcinoma (LIHC) cell lines. Based on the receiver operating characteristic (ROC) and survival analysis, SHCBP1 was considered as a potential diagnostic and prognostic biomarker. Furthermore, SHCBP1 expression was linked with tumor immunity and immunosuppressive microenvironment according to the correlation analysis of SHCBP1 expression with immune cells infiltration, immune checkpoint genes, and immune-related genes (MHC genes, chemokines, and chemokines receptors). Moreover, SHCBP1 expression correlated with tumor mutational burden (TMB), microsatellite instability (MSI), and neoantigens. The feature of SHCBP1 mutational landscape in pan-cancer was identified. Finally, we focused on investigating the clinical significance and the potential biological role of SHCBP1 in LUAD. Our study comprehensively uncovered that SHCBP1 could be identified as an immune-related biomarker for cancer diagnosis and prognosis, and a potential therapeutic target for tumor immunotherapy.

Isolation, identification and genetic characterization analysis of a fowl aviadenovirus serotype 4 strain from Tianjin, China.

A fowl aviadenovirus serotype 4 (FAdV-4), Y17215-1, was isolated from the liver of chickens with Hydropericardium-hepatitissyndrome(HHS) in a chicken farm of Tianjin, China. Obvious cytopathic effects were observed in the infected chicken liver hepatocellular carcinoma cell line (LMH cells) at 24h post infection (hpi), which consisted of enlarger and rounder shape of cells. The typical and specific green fluorescence was observed by indirect immunofluorescence assay (IFA). Tissue Culture Infectious Dose50 (TCID50) of it measured after five stable passage in LMH cells reached 106.5TCID50/0.1mL. The strain was inoculated through allantoic membrane of 10-day specific pathogen free(SPF) Chick embryos, the thicker allantoic membranes were observed at 120 hpi. 7-day-old SPF chickens were inoculated with the strain via intramuscular (i.m.) or intranasal (i.n.) injection which resulted in 100% mortality of test chickens. Additionally, the sickness and death of cohabitation chickens in the test group were observed which indicated that the virus can infect healthy chickens by horizontal transmission. The sick chickens showed depression, anorexia and diarrhea with green watery feces. Y17215-1-inoculated chickens mainly presented swollen liver with blood spot, and the enhancement of effusion or yellow gel like effusion that were observed in the pericardium through necropsy. Histopathological examination showed focal necrosis of hepatocytes and characteristic eosinophilic inclusion bodies in the cytoplasm. The results showed that the Y17215-1 isolate had high pathogenicity to SPF chickens. The phylogenetic analysis of the major structural proteins including hexon, fiber-1 and fiber-2 revealed that Y17215-1 strain belongs to C species of fowl aviadenovirus of aviadenovirus family, and has high homology with other Chinese strains isolated in recent years, but was distinct from ON1、MX-SHP95、KR5 and other foreign isolates. This study laid a foundation for further study of epidemiological investigation, pathogenic mechanism as well as the diagnosis and control technology of FAdV-4.

In Vitro Cytotoxicity and Oxidative Stress Evaluation of Valerian (Valeriana officinalis) Methanolic Extract in Hepg2 and Caco2 Cells

Traditional treatment methods are becoming popular and commonly used in many societies and have become the first treatment option for most people. While some of these methods are helpful, they can interact with medications the patient is taking for another disease and cause a variety of life-threatening risks. Valerian (catweed) plant is used in traditional medicine as a sleep aid due to its sedative effects. Valerian may also exert anticancer effect in vitro. In this study, the cytotoxicty and oxidative stress effects of valerian root extract were evaluated in human liver hepatocellular carcinoma (Hepg2) and human colorectal adenocarcinoma (Caco2) cell lines. The cytotoxicity was evaluated via the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide test. Total reactive oxygen species analysis was performed via a 2',7'-dichlorodihydrofluorescein diacetate assay in flow cytometry. Inhibition concentration 50 values were calculated as 936.6 and 1097.5 µg/mL in the Hepg2 and Caco2 cell lines, respectively. It was observed that valerian root extract did not induce oxidative stress in HepG2 and Caco2 cell lines. These results indicate that the use of valerian root extract as an alternative method in cancer treatment may not be effective and may cause a risk for public health. On the other hand, it may be safe at recommended tolerated concentrations since it does not cause oxidative stress.

Cytotoxicity and apoptosis response of hexagonal zinc oxide nanorods against human hepatocellular liver carcinoma cell line

Hexagonal zinc oxide nanorods (ZnO-NRs) were prepared via solution process using zinc nitrate hexahydrate (Zn(NO3)2·6H2O, 0.25 M), hexamethylenetetramine (HMT) and sodium hydroxide at low concentration (4 × 10-3 M in 100 ml of distilled water) and refluxed at 100 °C for an hour. The HMT was used because its act as a template for the nucleation and growth of zinc oxide nanorods. The X-ray diffraction patterns (XRD) clearly reveal that the grown product is pure zinc oxide. The diameters and lengths of the synthesized nanorods lie between 200 and 250 nm and 2–3 μm respectively as observed from the field emission scanning electron microscopy (FESEM). The chemical functional properties was analyzed by Fourier Transform infra-red (FTIR) spectroscopy. This study was designed to show the possible effect of ZnO-NRs in human liver cancer (HepG2) cells. It induces cytotoxicity via reactive oxygen species (ROS), mitochondrial membrane potential. For gene expression analysis, it induces apoptotic gene marker P53, Bax, caspase3, Bcl2 genes. Based on analysis and observations, it concludes that the ZnO-NRs were utilized for toxicity at dose dependent via ROS generation against human hepato cellular liver carcinoma cell line.

Evaluation of in-vitro bactericidal activity and anticancer effects of venom fractions of Pseudocerastes persicus snake

Introduction: It was reported that snake venom contains a wide mixture of proteins and peptides with various toxicological and pharmacological effects. In recent years, the isolation of effective molecules from snake venom as a drug to treatment of some incurable diseases is considered by many biological research centers. This investigation aimed to determine electrophoretic and chromatographic patterns of Pseudocerastes persicus snake venom and in-vitro study of bactericidal and anti-cancer properties of its fractions in a human hepatocellular liver carcinoma cell line (HepG2). Materials and methods: The fractions of venom were isolated and collected using preparative RP-HPLC. The Bactericidal activity of fractions in 20 μg/ml of protein concentration was investigated toward Gram-negative and Gram-positive bacteria. Antibacterial susceptibility tests were done using MTT and MIC assays. Furthermore, cell viability was measured by MTT reduction assay and confirmed with Neutral red uptake assay following exposure of HepG2 cancer cells to 20 and 40 μg/ml of protein concentrations of fractions having more antibacterial properties. Apoptotic effects were investigated using the comet assay. Results: SDS-PAGE pattern of the crude venom revealed 10 major bands with molecular weight ranging from 13.2 to 99.25 kDa and the single-step separation of different fractions from the venom by HPLC resulted in isolation and collection of 11 fractions. The results of this study showed that two fractions of snake venom have significant antibacterial activity against Gram-positive as compared to Gram-negative bacteria. On the other hand, our findings indicated that one of them is not toxic to the HepG2 cells. Discussion: We for the first time reported bactericidal and anticancer effects of isolated fractions of Persian horned viper venom. These properties make components of these fractions a favorable source for isolation of molecule(s) with antibacterial and antitumor activities.

Upregulation Mitochondrial Carrier 1 (MTCH1) Is Associated with Cell Proliferation, Invasion, and Migration of Liver Hepatocellular Carcinoma.

Among the primary causes of cancer-associated death in the world, liver hepatocellular carcinoma (LIHC) ranks the third. LIHC is defined as the sixth most frequently diagnosed carcinoma. The gene mitochondrial carrier 1 (MTCH1) is a protein-coding gene. Recent research suggests that MTCH1 may be associated with some diseases. Here, our study attempts to explore the role and implication of MTCH1 in LIHC. Kaplan Meier Plotter and GEPIA (Gene Expression Profiling Interactive Analysis) databases were employed to determine the expression of MTCH1 and its correlation with prognostic status in LIHC patients. For the first time, our results suggested that MTCH1 was aberrantly expressed in human pan-cancer and highly expressed in LIHC. Its high expression was closely associated with metastasis of tumor, stage of cancer, and poor survival of patients. Then, through enrichment analysis, MTCH1 was found to be closely related to RNA splicing in LIHC. Subsequently, we conducted a series of functional experiments. PCR data showed that LIHC cell lines and samples are highly expressed MTCH1. CCK-8 (Cell Counting Kit-8) assays and Transwell assays indicated that silencing MTCH1 certainly suppressed cell proliferation, migration, and invasion. These findings shed the clue that MTCH1 could be regarded as the potential prognosis biomarker of LIHC and a promising therapeutic target for LIHC.

4N donor atoms moiety of transition metal complexes of a Schiff base ligand: Synthesis, characterization and biological activities study

Prepared new Schiff base ligand [H2L] with 4N donor atoms moiety by reacting amine derivative of o-phenylenediamine hydrazide moiety with benzaldehyde. Co(II), Ni(II), Cu(II) and Zn(II) of the new Schiff base were prepared by using a molar ratio of ligand: metal as (1:1). The structure of the Schiff base and the prepared complex was determined by 1H NMR, UV–Vis, IR, Mass spectrometry and molar conductivity. The thermal stability of the complexes had been studied by TGA. The spectral statement displays that the ligand behaves as bidentate ligand. On the basis of the spectral studies, reveal that an octahedral geometry had been assigned for all complexes. The Anti-proliferative effect was screened against human hepatocellular liver carcinoma cell line (HepG2) and antimicrobial activity had been inspected. Result points that complexes demonstrate the increase cytotoxicity in proliferation to cell lines as compared to free ligand.

Efficient synthesis and molecular docking studies of new pyrimidine-chromeno hybrid derivatives as potential antiproliferative agents

Various novel heterocyclic compounds containing pyrimidine nuclei 5H-chromeno[4,3-d]pyrimidine (4a–c, e–h, l–r, t) and pyrimidine-5-yl-(2-hydroxyphenyl)methanone (5a, c, d, f–k, m–o, r, s, u) were synthesized from the reaction of guanylhydrazones (2a–u) and 3-formylchromone (3). These compounds were tested against human liver hepatocellular carcinoma cell line (HepG2) and human breast adenocarcinoma cell line (MDA-MB-231) using the MTT assay method. Furthermore, molecular docking calculations were performed to compare the biological activities of various novel heterocyclic compounds against cancer proteins. In these calculations, the protein used are crystal structure of the BRCT repeat region from the breast cancer associated protein, 1JNX, crystal structure of VEGFR kinase (liver cancer) protein, 3WZE, and crystal structure of an allosteric Eya2 phosphates inhibitor (lung cancer) protein, 5ZMA, respectively. After molecular docking calculations, absorption, distribution, metabolism, and excretion/toxicity analysis was performed to examine the properties of various novel heterocyclic compounds for their future use as drugs.

Nucleotide analogues containing a pyrrolidine, piperidine or piperazine ring: Synthesis and evaluation of inhibition of plasmodial and human 6-oxopurine phosphoribosyltransferases and invitro antimalarial activity.

Parasites of the Plasmodium genus are unable to produce purine nucleotides de novo and depend completely on the salvage pathway. This fact makes plasmodial hypoxanthine-guanine-(xanthine) phosphoribosyltransferase [HG(X)PRT] a valuable target for development of antimalarial agents. A series of nucleotide analogues was designed, synthesized and evaluated as potential inhibitors of Plasmodium falciparum HGXPRT, P.vivax HGPRT and human HGPRT. These novel nucleoside phosphonates have a pyrrolidine, piperidine or piperazine ring incorporated into the linker connecting the purine base to a phosphonate group(s) and exhibited a broad range of Ki values between 0.15 and 72μM. The corresponding phosphoramidate prodrugs, able to cross cell membranes, have been synthesized and evaluated in a P.falciparum infected human erythrocyte assay. Of the eight prodrugs evaluated seven exhibited invitro antimalarial activity with IC50 values within the range of 2.5-12.1μM. The bis-phosphoramidate prodrug 13a with a mean (SD) IC50 of 2.5±0.7μM against the chloroquine-resistant P.falciparum W2 strain exhibited low cytotoxicity in the human hepatocellular liver carcinoma (HepG2) and normal human dermal fibroblasts (NHDF) cell lines at a concentration of 100μM suggesting good selectivity for further structure-activity relationship investigations.

Protective effect of Zingiber zerumbet Smith extract on thermotolerance

Background: Global warming causes severe heat conditions. Heat stress contributes to higher morbidity of heatstroke in human and mortality in livestock. To protect them from heat stress, thermotolerance mechanisms were widely studied, and some studies suggest relationship between heat shock proteins (HSPs) and thermotolerance. HSPs were not induced by only heat shock but also some stimulations including bioactive compounds from plants. Zingiber zerumbet is a perennial herb found in many tropical countries, including Thailand. The rhizome of Zingiber zerumbet contains zerumbone that is a bioactive compound to induce HSPs expression in animal cells.Objective: To prevent higher morbidity of heatstroke in human and mortality in livestock by the heat stress, we investigated the effect of zerumbone, the extract of Zingiber zerumbet Smith, on thermotolerance, using a cell line and mice.Methods: The murine liver hepatoma cell line, Hepa1c1c7 cells, were incubated in medium supplemented with extract from rhizome of Zingiber zerumbet Simith containing zerumbone, and then the expression of heat shock proteins (HSP) 40, 70 and 90 were investigated by western blotting. Furthermore, we established the evaluation system of thermotolerance using mice, and studied the effect of the extract on the growth rate of mice under the heat shock treatment. Briefly, 4 weeks old C57BL6 mice were fed that with the extract (or vehicle) for a week before the first heat shock treatment (38 °C for an hour). Before and after five days heat treatment, body weights were measured. The protein expressions of heat shock proteins in liver were measured by western blotting using HSPs antibodies.Results: The extract of Zingiber zerumbet rhizome, equivalent to 50 μM zerumbone, significantly increased the expression of heat shock proteins (HSP40, HSP70, HSP90). The growth rate of the mice under the heat treatment were lower than control. The feeding with the extract containing 25 ppm zerumbone have significantly attenuated the decline of the growth rate led by the heat treatment, whereas there was little effect on mouse growth rate grown under normal conditions. The protein expression of HSP70 in the liver of zerumbone-fed mice was upregulated compared with control mice, equivalent to heat treatment without zerumbone. On the other hand, both treatments of zerumbone and heat resulted in highest HSP70 expression among four groups.Conclusion: Our study demonstrated that oral administration of the extract of Zingiber zerumbet Smith led to the attenuation of decline of growth rate induced by heat treatment. HSP70 expression in murine liver was enhanced by either feeding the extract or heat treatment. More interestingly, HSP70 expression was further enhanced by both treatments of zerumbone and heat. These results suggested that zerumbone may contribute to thermotolerance via, at least, HSP70 expression.Keywords: Zingiber zerumbet, thermotolerance, heat shock protein

Crude venom of Pseudocerastes persicus snake: From the antibacterial to anticancer effects

Antibiotic resistance has been reported as one of the world's most critical public health problems. Recent investigations have demonstrated that the venom of some species of snakes has antimicrobial and anticancer activities. The purpose of this study was to examine the antibacterial activity of Persian horned viper venom toward Escherichia coli, Bacillus subtilis, and Staphylococcus aureus bacteria and also investigate antitumor effects of concentrations of venom with higher antibacterial activity in a human hepatocellular liver carcinoma cell line (HepG2) to evaluate the potential use of its components as antimicrobial and anticancer agents. Bactericidal activity of crude venom in concentrations of 6.25-400 μg/ml was performed using MTT reduction, minimum inhibitory concentration (MIC), agarwell diffusion, and disc diffusion methods. Tetracycline (50 μg/ml) was used as a standard antibiotic. Cytotoxic effect in HepG2 cells was measured by MTT reduction assay and confirmed with neutral uptake assay following exposure of cells with different concentrations of venom (50-400 μg/ml). The apoptotic effect was investigated using the comet assay. Our findings demonstrated that venom displays higher inhibitory effects against Gram-positive bacteria as compared to Gramnegative. Furthermore, venom showed anticancer activity on the HepG2 cell line through induction of apoptosis and necrosis. The results of this study confirmed that the venom of Persian horned viper induces antibacterial and anticancer effects. These properties make the venom of this viper a potential source for isolation of effective molecule(s) having antibacterial and antitumor activity.

Synthesis, characterization, DNA binding ability, in vitro cytotoxicity, electrochemical properties and theoretical studies of copper(II) carboxylate complexes

Four copper(II) carboxylate complexes, namely [Cu2(μ-3,5-dinitrobenzoate-O,O')4(4-(dimethylamino)benzaldehyde)2] (1), [Cu2(μ-benzoate-O,O')4(benzoic acid)2] (2), [Cu2(μ-benzoate-O,O')4(H2O)2] [Cu(benzoate-O,O')2(imz)2].2H2O (3) and [Cu(benzoate-O,O')2(2-Me-imz)2] (4) (imz = imidazole, 2-Me-imz = 2-methyl-imidazole), were synthesized and comprehensively characterized by elemental analysis, spectroscopic methods, single crystal X-ray diffraction, cyclic voltammetry (CV), topological analysis as well as theoretical studies. Single crystal X-ray diffraction revealed that products 1 and 2 are dinuclear paddle-wheel complexes, compound 3 is a co-crystal containing mononuclear and dinuclear blocks, while compound 4 is a mononuclear complex. Hirshfeld surface analysis of the compounds rationalized different types of hydrogen bonds, which also lead to the generation of H-bonded networks in 3 and 4. Their topological analysis disclosed a uninodal 4-connected 2D layer with sql topology in 3 and a uninodal 2-connected 1D chain with 2C1 topology in 4. The interaction of 1–4 with calf thymus DNA was investigated by UV–visible and fluorescence spectroscopy, revealing a moderately strong non-intercalative mode of interaction. In vitro cytotoxicity study of the complexes on HepG2 (human liver hepatocellular carcinoma) cell lines revealed a significant inhibition activity. Electrochemical study of the complexes in CH3CN (1–3) and DMSO (4) solution showed a one electron transfer corresponding to Cu(III)Cu(II)/Cu(II)Cu(II) and Cu(II)Cu(II)/Cu(I)Cu(II) redox couples. The ΔE and Ipa/Ipc values suggest that the redox couples are quasireversible. The DFT study was performed to further rationalize the crystal structures and nuclearity of the obtained copper(II) complexes.

Transferrin receptor regulates malignancies and the stemness of hepatocellular carcinoma-derived cancer stem-like cells by affecting iron accumulation

Iron metabolism is essential because it plays regulatory roles in various physiological and pathological processes. Disorders of iron metabolism balance are related to various cancers, including hepatocellular carcinoma. Cancer stem-like cells (CSCs) exert critical effects on chemotherapy failure, cancer metastasis, and subsequent disease recurrence and relapse. However, little is known about how iron metabolism affects liver CSCs. Here, we investigated the expression of transferrin receptor 1 (TFR1) and ferroportin (FPN), two iron importers, and an upstream regulator, iron regulatory protein 2 (IRP2), in liver hepatocellular carcinoma (LIHC) and related CSCs. The expression levels of TFR1, FPN and IRP2 were analysed using the GEPIA database. CSCs were derived from parental LIHC cells cultured in serum-free medium. After TFR1 knockdown, ROS accumulation and malignant behaviours were measured. The CCK-8 assay was performed to detect cell viability after TFR1 knockdown and erastin treatment. TFR1 expression was upregulated in LIHC tissue and CSCs derived from LIHC cell lines, prompting us to investigate the roles of TFR1 in regulating CSCs. Knockdown of TFR1 expression decreased iron accumulation and inhibited malignant behaviour. Knockdown of TFR1 expression decreased reactive oxygen species (ROS) accumulation induced by erastin treatment and maintained mitochondrial function, indicating that TFR1 is critical in regulating erastin-induced cell death in CSCs. Additionally, knockdown of TFR1 expression decreased sphere formation by decreasing iron accumulation in CSCs, indicating a potential role for TFR1 in maintaining stemness. These findings, which revealed TFR1 as a critical regulator of LIHC CSCs in malignant behaviour and stemness that functions by regulating iron accumulation, may have implications to improve therapeutic approaches.

An Efficient Procedure of Synthesis Acyclic C-Glycosides of Thiazolo [4, 5-b]Pyrazine and Imidazo[4,5-d]Thiazole with Expected Anti-Cancer Activities

In this research, we aimed to synthesize a new series of C-glycosides that have thiazole-4, 5-diamine base. C-glycosides 8 was prepared by coupling compound 7 with D-glucose in the presence of iodine used as an oxidant/promoter dissolved in acetic acid and stirring at room temperature. Compound 8 was protected by reaction with acetic anhydride in the presence of pyridine gave compound 9. Furthermore, cyclization compound 7 with hydrazine hydrate and D-glucose gave the cyclic glycosides analogs 10. The compound 7 was condensed with phenyl hydrazine hydrochloride and D-xylose gave C-glycoside 11. The anticancer activity of the newly synthesized compounds was tested in vitro for their anticancer activities against human colorectal carcinoma (HCT-116), human prostate adenocarcinoma (PC-3), and human liver hepatocellular carcinoma (HepG-2) cell lines.

Novel Harmicines with Improved Potency against Plasmodium

Harmicines represent hybrid compounds composed of β-carboline alkaloid harmine and cinnamic acid derivatives (CADs). In this paper we report the synthesis of amide-type harmicines and the evaluation of their biological activity. N-harmicines 5a–f and O-harmicines 6a–h were prepared by a straightforward synthetic procedure, from harmine-based amines and CADs using standard coupling conditions, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIEA). Amide-type harmicines exerted remarkable activity against the erythrocytic stage of P. falciparum, in low submicromolar concentrations, which was significantly more pronounced compared to their antiplasmodial activity against the hepatic stages of P. berghei. Furthermore, a cytotoxicity assay against the human liver hepatocellular carcinoma cell line (HepG2) revealed favorable selectivity indices of the most active harmicines. Molecular dynamics simulations demonstrated the binding of ligands within the ATP binding site of PfHsp90, while the calculated binding free energies confirmed higher activity of N-harmicines 5 over their O-substituted analogues 6. Amino acids predominantly affecting the binding were identified, which provided guidelines for the further derivatization of the harmine framework towards more efficient agents.

DFT, anticancer, antioxidant and molecular docking investigations of some ternary Ni(II) complexes with 2-[(E)-[4-(dimethylamino)phenyl]methyleneamino]phenol

The biological activities of some ternary nickel complexes with a Schiff base obtained from 4-dimethylaminobenzaldehyde and 2-aminophenol have been reported. The Schiff base (HL1) acts as a primary ligand whereas, anthranilic acid (HL2), 2-nitroaniline (HL3), alanine (HL4) and histidine (HL5) act as secondary ligand or co-ligand. The anticancer activity of these compounds was studied against human colon carcinoma (HCT-116), human hepatocellular liver carcinoma (HEPG-2) and human breast carcinoma (MCF-7) cell lines. As per the results, the compounds were active against the cell lines. The antioxidant activity of the same compounds was evaluated using DPPH (1,1-diphenyl-2-picryl-hydrazyl) radical scavenging and compared with ascorbic acid. The DFT computations for these compounds were made to understand the bonding mode by a GAUSSIAN 09 program. Moreover, a docking analysis using Autodock 4.2 software package was carried out against the tyrosine kinase receptor (PDB ID: 1M17). In addition, QSAR investigation was also performed to understand the biological potency of the ligand.

HEPATOPROTECTIVE ACTIVITY OF ETHANOLIC STEM BARK EXTRACT OF KNEMA ATTENUATA (HOO K. F. AND THOMSON) WARB

Objective: To investigate the hepatoprotective activity of ethanolic stem bark extract (ESBE) of Knema attenuata against carbon tetrachloride (CCl4) induced hepatotoxicity in Wistar rats using both in vivo and in vitro models.
 Methods: Animals were treated orally with ESBE (250 mg kg-1 and 500 mg kg-1) once daily for 6 d and CCl4 on the 4th d. On the 7th d, animals were sacrificed and the blood samples were collected to measure the serum levels of biochemical parameters, whereas the liver homogenates were utilized for estimating the antioxidant defense. The hepatoprotective efficacy of the extract was further ensured in vitro using human liver hepatocellular carcinoma (HepG2) cell line against CCl4 induced toxicity. The cell line viability was determined using 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.
 Results: ESBE effectively reduced (p<0.001) the elevated serum levels of Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), and Alkaline phosphatase (ALP) when compared to the toxicant control group. ESBE 500 mg kg-1significantly raised the antioxidant defense (p<0.0001) by reducing the malondialdehyde (MDA) level and enhancing hepatic reduced glutathione (GSH) level in comparison to the CCl4 control group. The in vitro effect was investigated using CCl4 exposed HepG2 cells. Pretreatment with ESBE showed a dose-dependent increase in percentage cell viability ranged between 44 to 57% at 12.5-100 μg ml-1concentrations (p<0.001, when compared to the control cells). 
 Conclusion: Present study confirms the hepatoprotective activity of the stem bark extract of K. attenuata against CCl4‑induced liver damage.