Hepatic Cancer Cell Line HepG2 Research Articles

Bisimidazolium Salt Glycosyltransferase Inhibitors Suppress Hepatocellular Carcinoma Progression In Vitro and In Vivo.

Hepatocellular carcinoma is a leading cause of cancer death, and the disease progression has been related to glycophenotype modifications. Previously synthesized bisimidazolium salts (C20 and C22) have been shown to selectively inhibit the activity of glycosyltransferases in cultured cancer cell homogenates. The current study investigated the anticancer effects of C20/C22 and the possible pathways through which these effects are achieved. The therapeutic value of C20/C22 in terms of inhibiting cancer cell proliferation, metastasis, and angiogenesis, as well as inducing apoptosis, were examined with hepatic cancer cell line HepG2 and a xenograft mouse model. C20/C22 treatment downregulated the synthesis of SLex and Ley sugar epitopes and suppressed selectin-mediated cancer cell metastasis. C20/C22 inhibited HepG2 proliferation, induced cell-cycle arrest, increased intracellular ROS level, led to ER stress, and eventually induced apoptosis through the intrinsic pathway. Furthermore, C20/C22 upregulated the expressions of death receptors DR4 and DR5, substantially increasing the sensitivity of HepG2 to TRAIL-triggered apoptosis. In vivo, C20/C22 effectively inhibited tumor growth and angiogenesis in the xenograft mouse model without adverse effects on major organs. In summary, C20 and C22 are new promising anti-hepatic cancer agents with multiple mechanisms in controlling cancer cell growth, metastasis, and apoptosis, and they merit further development into anticancer drugs.

Some Efficient Synthetic Approaches to Construct Heterocyclic Systems: Thesis Abstract

Heterocyclic compounds containing nitrogen, oxygen and sulfur atoms possess various interesting biological activities. Among these compounds, 1,3-oxathiolan-5-one and its diverse derivatives which are literature documented ones of the most potent core structures in drug design and pharmaceutical industry. This thesis describes the utility of 1,3-oxathiolan-5-one in the synthesis of pyridinonethiol and their annulated systems through multicomponent reaction of 2-methyl-2-phenyl-1,3-oxathiolan-5-one (1), aniline and α,β-unsaturated carbonyl compounds. On the other hand, different bioactive heterocyclic systems such as thiophene, furo-oxathiolone, mercapto-diphenylcyclopenta-2,4-dien-one, triazole and thiadiazine have been efficiently synthesized via tandem reactions of 1,3-oxathiolan-5-one 1 with different nucleophilic reagents. The structures of newly synthesized compounds were confirmed by spectral data, elemental analyses and mechanically discussed. Also, in vitro antioxidant and antitumor activities of these compounds were evaluated. In addition, the synthesized compounds were screened against hepatic cancer cell line HepG-2, as the optimistic results exhibited that compounds 29, 33 and 34 have excellent antioxidant and anti HepG-2 activities.

The role of membrane-bound metal ions in toxicity of a human cancer cell-active pore-forming toxin Cry41Aa from Bacillus thuringiensis

Bacillus thuringiensis crystal (Cry) proteins, used for decades as insecticidal toxins, are well known to be toxic to certain insects, but not to mammals. A novel group of Cry toxins called parasporins possess a strong cytocidal activity against some human cancer cells. Cry41Aa, or parasporin3, closely resembles commercially used insecticidal toxins and yet is toxic to the human hepatic cancer cell line HepG2, disrupting membranes of susceptible cells, similar to its insecticidal counterparts. In this study, we explore the protective effect that the common divalent metal chelator EGTA exerts on Cry41Aa's activity on HepG2 cells. Our results indicate that rather than interfering with a signalling pathway as a result of chelating cations in the medium, the chelator prevented the toxin's interaction with the membrane, and thus the subsequent steps of membrane damage and p38 phosphorylation, by removing cations bound to plasma membrane components. BAPTA and DTPA also inhibited Cry41Aa toxicity but at higher concentrations. We also show for the first time that Cry41Aa induces pore formation in planar lipid bilayers. This activity is not altered by EGTA, consistent with a biological context of chelation. Salt supplementation assays identified Ca2+, Mn2+ and Zn2+ as being able to reinstate Cry41Aa activity. Our data suggest the existence of one or more metal cation-dependent receptors in the Cry41Aa mechanism of action.

Lycium barbarum polysaccharides grafted with doxorubicin: An efficient pH-responsive anticancer drug delivery system

Lycium barbarum polysaccharide-doxorubicin (LBP-DOX) nanoparticles were synthesized via a one-step Schiff base reaction between oxidized Lycium barbarum polysaccharide (oxidized LBP) and doxorubicin (DOX). The structure of LBP-DOX nanoparticles was characterized by multi-spectral methods, 1H nuclear magnetic resonance (1H NMR), transmission electron microscopy (TEM), and thermo gravimetric analysis (TGA). The rheological properties of LBP-DOX nanoparticles were studied. With the increase of shear rate, the viscosity of LBP and LBP-DOX nanoparticles gradually decreases at 25 °C. Under the same strain, the storage modulus (G′) and loss modulus (G″) of LBP-DOX nanoparticles were inferior to LBP. The DOX release from LBP-DOX nanoparticles was pH-dependent and was accelerated by decreasing pH. Cytotoxicity study showed that the LBP-DOX nanoparticles have significantly enhanced cytotoxicity in vitro, especially for human Hepatic cancer cell line HepG2.

In Vitro Cytotoxic and Antiproliferative Activity of Cydonia oblonga flower petals, leaf and fruit pellet ethanolic extracts. Docking simulation of the active flavonoids on anti-apoptotic protein Bcl-2

AbstractThis study aimed to compare in vitro cell cytotoxicity and antiproliferative potency of three standardized ethanolic extracts (5mg GAE/mL sample) from quince flower petals, leaves and fruit pellet on four cell lines (L-929, and HepG2, Caco-2 and BT-20 respectively). Comparative analytical qualitative studies (HPTLC) indicated that if quince leaf extracts (Col40) mainly contain quercetin and kaempferol derivates, the flower petal extracts (Cof40) contain caffeoylquinic acid derivates, while the fruit pellet extracts (Cop40) are comprised of quercetin and caffeoylquinic acid derivates. Pharmacological studies demonstrated the lack of toxicity of test extracts; the most important antiproliferative effects were observed on the hepatic cancer cell line HepG2 (up to 75%, 53% and 70% inhibition in the case of Col40, Cof40 and Cop40 test extracts), followed by the colon cancer cell line Caco-2 (up to 69%, 77% and 40% inhibition) and breast cancer cell line BT-20 (up to 54%, 61% and 19% inhibition). The docking simulations on hyperoside, isoquercitrin, astragalin, and quercetin and kaempferol compared to the synthetic co-crystallized LI0 A1000 ligand (a strong inhibitor of anti-apoptotic protein Bcl-2) indicated astragalin as the most feasible protein inhibitor, but quercetin and kaempferol respected all the parameters involved in the Lipinski rule, making them the most promising antiproliferative candidates.

Abstract 3490: Mechanism of action for antioxidant Twendee X on hepatic cancer cell line HepG2

Abstract [Purposes of the study] Reactive oxygen species (ROS) are associated with clinical condition of cancer patient, and closely involved in cancer growth and metastasis. Twendee X (TWX) is a composition consisting of Vitamin C, Glutamine, Cystine, Riboflavin, Succinic acid, Fumaric acid, Coenzyme Q10, and Niacin. TWX strongly reduces oxygen radicals or ROS in vitro and in vivo (World Intellectual Property Organization WO2013/072441 A1 COMPOSITION FOR PROTECTION AGAINST CELL-DAMAGING EFFECTS). TWX has significant effects of reducing tumor growth and inhibition metastasis. Hydrogen peroxide level (d-ROMs test) of tumor bearing nude mouse in serum was significantly reduced and increased NK-activity (2016 AACR meeting). Mechanism of action was examined for TWX using hepatic cancer cell line HepG2. [Experimental procedures] Human hepatic cancer cell line HepG2 was used in this study. Cell proliferation experiments of antioxidant by ICDD (France). Metabolome analysis was by Human Metabolome Technologies (Tokyo). In both experiments, HepG2 cell line treated one hour with TWX (60ug/ml). [Data] TWX inhibits the RAF-MEK-ERK pathway starting from ERK. Metabolome analysis shows total shift of metabolism, and especially increase mitochondrial metabolism and ATP production. [Conclusion] Effects of TWX on cancer cell line are not only increase immune activity, but also inhibit proliferation pathway, and mitochondrial metabolism change. Several publications using Vitamin-E or Cysteine as anti-oxidant showed cancer progression or increase metastasis. However, Vitamin-E or Cysteine has limited anti-oxidant effects and hydrogen peroxide level (d-ROMS test) dose not change. On the other hand, TWX has significant effects reduce hydrogen peroxide level, and Japanese Association for Dementia Prevention already started clinical trial for MCI and after stroke patients. For the cancer patients, clinical trial using TWX combination with standard therapy will be start soon. Citation Format: Haruhiko Inufusa. Mechanism of action for antioxidant Twendee X on hepatic cancer cell line HepG2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3490.

Cross-linked hyaluronic acid gel inhibits metastasis and growth of gastric and hepatic cancer cells: in vitro and in vivo studies.

Cross-linked hyaluronic acid gel (CHAG) has been used to prevent postoperative adhesion of abdominal tumorectomy. However, its effect on tumor cells is still unknown. This paper was designed to investigate the effect of CHAG on metastasis and growth of tumor cells. Migration and invasion assays, Western blotting, pull down assay, siRNA interference, and nude mice implantation tumor model were applied in this study. The results of in vitro experiments with gastric cancer cell line AGS and hepatic cancer cell line HepG2 showed that CHAG inhibited the migration and invasion activities, the MAPK and PI3K/Akt mediated signaling, the activation of small G proteins Rac1 and RhoA, and the expression of MMPs and PCNA initiated by EGF, through blocking the activation of EGFR. CHAG also had inhibitory effect on activation of other membrane receptors, including integrin and VEGFR. When the expression of hyaluronic acid receptors (CD44 or RHAMM) was interfered, the above inhibitory effects of CHAG still existed. In vivo experimental results showed that CHAG suppressed colonization, growth and metastasis of gastric cancer cell line SGC-7901 in peritoneal cavity of nude mice. In conclusion, CHAG had inhibitory effect on tumor cells, through covering cell surface and blocking the interaction between extracellular stimulative factors and their receptors.

Two new metabolites from a soil fungus Curvularia affinis strain HS-FG-196

Two new metabolites, pyrenocine J (1) and pyrenochaetic acid D (2), together with two known metabolites, pyrenocine A (3) and pyrenochaetic acid A (4), were isolated from a soil fungus, Curvularia affinis strain HS-FG-196. Their structures were established by extensive spectroscopic analysis. Compound 1 showed cytotoxic activity against the human hepatic cancer cell line HepG2 with an IC50 value of 28.5 μg/ml.

Biliary epithelial cells inhibit proliferation of co-cultured hepatic cancer cell line HepG2

Objective To investigate the possible effect of mouse intrahepatic biliary epithelial cell line(mIBEC) on co-cultured human hepatoma cell lines HepG2.Methods HepG2 and mIBEC cells were co-cultured in a membrane-separated Transwell system.CellTiter 96 ○R AQueous One Solution Cell Proliferation Assay(Promega Corporation,Madison,WI) was used to examine HepG2 cell proliferation in the system with or without co-cultured mIBEC.Real-time PCR(RT-PCR) was used to determine Ki67 and caspase3 mRNA expression in HepG2 cells in a system with or without co-cultured mIBEC.Western blotting analysis was used to determine caspase3 protein level in HepG2 cells.Results The proliferation of the co-cultured HepG2 cells was significantly lower than those cultured alone(P 0.01).Expression of Ki67,a cell proliferation marker,was also significantly down-regulated in mIBEC co-cultured HepG2 cells(P 0.05).The levels of caspase3 mRNA and protein were significantly up-regulated in mIBEC co-cultured HepG2 cells compared with HepG2 cells cultured alone(P 0.05).Conclusion mIBEC can inhibit the proliferation of co-cultured HepG2,and caspase3 activation might be one of the reasons for the inhibitory effect of mIBEC against HepG2 cells.

Abstract 1055: Character of tumor assocated protien recognized by monoclonal antibody against yunnan gejiu lung cancer

Abstract Character of tumor assocated protien recognized by monoclonal antibody against Yunnan Gejiu lung cancer: about protein N35 WANG Qinqin The Clinical Reseach Center. The First Hospital of Kunming Medial College Kunming, Yunnan, P.R. China 650032 ABSTRACT Objectives: To identify and characterize lung cancer associated protein N35 and attempt to learn the prospective possibility of the clinical application of the protein N35. Methods: Immunoprecipitation, immunoblotting, differential centrifigation and subcellular assay, immunohistochemistry, Nglycanase digestion, mitotic cell immunoflourescence and multiple methods of affinity chromatography have been used with the monoclonal antibody N-35 to detect the distribution of the protein N35 among the various cancer cell lines and normal human tissue, the relationship between the protein N35 and glycoprotein, the location of the subcellular structure and chromosomal domain of the protein N35,the most effective way of purification of tumor associated protein N35. Results: The protein N35 is a glycoprotein, distributes to the human lung cancer cell line GLC-82, human cervical cancer cell line Hela, human hepatic cancer cell line HepG-2 and human breast cancer cell line PMC with different relative molecular mass(Mr), but no expression of the protein ingredient in normal human fresh tissue; concentrates at the nuclei significantly, much more than at the mitochondrail and membrane, locates at the centriole of the chromosomal domain. Conclusions: The lung cancer associated protein N35 might be expressed only by the cancer cells and related with the proliferation of cancer cells as a role of tumor cell growth regulator. Keywords: Lung cancer cells, Tumor associated protein, Glycoprotein, Centriole Qinqin Wang Prof of The Clinical Reseach Center. The First Hospital of Kunming Medial College 295 Xichang Rd. 650032 Kunming Yunnan, P.R. China Tel:868714191985 Fax: 868715336015 Email: zys@public.km.yn.cn Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1055.

The effect of fatty acid-CoA ligase 4 on the growth of hepatic cancer cells

In this study, we detected the expression of FACL4 mRNA in 40 patients with hepatic carcinoma and its adjacent normal tissues by semi-quantitative RT-PCR. The changes of proliferation and apoptosis of hepatic cancer cell line HepG2 with FACL4 protein expression were examined by MTT and flow cytometry respectively after FACL4 selective inhibitor triacsin C treatment. The activity related to apoptosis of proteinases, caspase-3, caspase-8 and caspase-9, were detected by colorimetry. The expression related to apoptosis of protein, wt-p53，Bax, and Bcl-2, in HepG2 cells were evaluated by S-P immunocytochemical dyeing. The results were: (1) FACL4 mRNA was expressed in 95.0% of hepatic cancer tissue, while the positive expression of FACL4 mRNA was 82.5% in cancer adjacent normal liver tissues. Moreover, there was a statistically significant increased in quantity of FACL4 mRNA in cancer tissues compared with adjacent normal liver tissues. (2) The concentration of triacsin C（0.5-2mg/L）could inhibit the proliferation and induce the apoptosis of HepG2 cells significantly in a dose- and time-effect. (3) During the apoptosis of HepG2 cells induced by triacsin C, flow cytometry coupled with Rhodamine 123 dyeing showed that mitochiondrial transmembrane potential of HepG2 declined significantly，and the activity of caspase-9 and caspase-3 increased more remarkably than caspase-8. Besides, the increased apoptosis was accompanied by increased Bax, and decreased wt-p53 and Bcl-2 protein levels. The present study suggested that FACL4 might play a role in the growth of hepatic cancer cells. FACL4 selective inhibitor triacsin C leads to a marked growth inhibition of human liver tumor cells, based on the inhibition of proliferation and induction of apoptosis. The apoptotic process was mediated by intrinsic mitochondrial apoptotic pathway due to activation of caspase-9 and caspase-3. The increased apoptosis was accompanied by upregulation of Bax, and decreased wt-p53 and Bcl-2 protein level.

Intercellular imaging by a polyarginine derived cell penetrating peptide labeled magnetic resonance contrast agent, diethylenetriamine pentaacetic acid gadolinium

The cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro. Fluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT). The molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group. The newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.

Physicochemical properties and antitumor activities of water-soluble native and sulfated hyperbranched mushroom polysaccharides

A water-soluble hyperbranched β-glucan, coded as TM3b, extracted from sclerotia of an edible fungus ( Pleurotus tuber-regium) was fractioned into eight fractions coded as F1–F8 by a nonsolvent addition method. Five fractions were treated with chlorosulfonic acid at 35 °C to synthesize successfully sulfated derivatives coded as S-F2, S-F3, S-F4, S-F5, and S-F8 with degree of substitution of 0.28–0.54. The 13C NMR results of these sulfated β-glucans indicated that while the C-6 position was fully substituted, C-2, C-3, and C-4 were only partially substituted by the sulfate groups. The weight-average molecular weights ( M w) and intrinsic viscosities ([ η]) of the native and sulfated TM3b fractions were determined using multi-angle laser light scattering and viscometry in 0.15 M aq NaCl at 25 °C, respectively. The dependences of [ η] on M w for TM3b and sulfated TM3b were found to be [ η ] = 0.18 M w 0.28 ± 0.03 ( M w range from 3.30 × 10 4 to 3.90 × 10 7) and [ η ] = 2.24 × 10 - 2 M w 0.52 ± 0.06 ( M w range from 3.24 × 10 4 to 3.15 × 10 5) in 0.15 M aq NaCl at 25 °C, respectively. It revealed that both the native TM3b and its sulfated derivatives exist in a spherical chain conformation in 0.15 M aq NaCl. Furthermore, the native and sulfated TM3b fractions showed potent antitumor activities in vivo and in vitro. The sulfated derivatives exhibited relatively higher in vitro antitumor activity against human hepatic cancer cell line HepG2 than the native TM3b. Water solubility and introduction of sulfate groups were the main factors in enhancing the antitumor activities.

Character of tumor associated protein recognized by monoclonal antibody against Yunnan gejiu lung cancer

Objectives: To identify and characterize lung cancer associated protein N35 and attempt to learn the prospective possibility of the clinical application of the protein N35. Methods: Immunoprecipitation, immunoblotting, differential centrifigation and subcellular assay, immunohistochemistry, N-glycanase digestion, mitotic cell immunoflourescence and multiple methods of affinity chromatography have been used with the monoclonal antibody N-35 to detect the distribution of the protein N35 among the various cancer cell lines and normal human tissue, the relationship between the protein N3S and glycoprotein, the location of the subcellular structure and chromosomal domain of the protein N35,the most effective way of purification of tumor associated protein N35. Results: The protein N35 is a glycoprotein, distributes to the human lung cancer cell line GLC-82, human cervical cancer cell line Hela, human hepatic cancer cell line HepG-2 and human breast cancer cell line PMC with different relative molecular mass(Mr), but no expression of the protein ingredient in normal human fresh tissue; concentrates at the nuclei significantly ,much more than at the mitochondrail and membrane, locates at the centriole of the chromosomal domain. Conclusions: The lung cancer associated protein N35 might be expressed only by the cancer cells and related with the proliferation of cancer cells as a role of tumor cell growth regulator.