betadex and Carcinoma--Hepatocellular

Topics: Antineoplastic Agents; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line, Tumor; Cholic Acid; Doxorubicin; Graphite; Humans; Hyaluronic Acid; Hyperthermia, Induced; Liver Neoplasms; Pharmaceutical Preparations; Phototherapy; Polymers

Cholesterol, the principal sterol in mammalian cells, has been reported to play a role in the pathogenesis of several diseases through autophagy. Due to its insoluble characteristic, all in vitro cholesterol experiments are performed using dimethyl sulphoxide, methyl-β-cyclodextrin, and ethanol co-solvents. To investigate whether the types of solvents have different effects on cholesterol-induced cell behaviors, we analyzed the effects and mechanisms of autophagy induced by solubilized-cholesterol in hepatic cells. We found that both solubilized-cholesterol and involved solvents could induce autophagy. Solubilized-cholesterol could further enhance the LC3-II expression with or without the pre-treatment with lysosomal blockers compared with the single-solvent groups, indicating that cholesterol could sensitize cells to solvents-induced autophagy. Besides, solubilized-cholesterol and single-solvent treatment could repress the activation of AKT-mTOR pathway. Furthermore, cholesterol solubilized in methyl-β-cyclodextrin could induce apoptosis while other solubilized-cholesterol or single solvent groups could not, suggesting that different dissolve methods may affect the cytotoxic of cholesterol. These results strongly suggest that the effect of solvent should be taken into consideration in further in vitro cholesterol studies.

Topics: Animals; Apoptosis; Autophagy; beta-Cyclodextrins; Carcinoma, Hepatocellular; Caspase 3; Cell Line, Tumor; Cell Survival; Cholesterol; Hepatocytes; Liver Neoplasms; Lysosomes; Microscopy, Electron, Transmission; Microtubule-Associated Proteins; Proto-Oncogene Proteins c-akt; Rats; Solvents; TOR Serine-Threonine Kinases

β-CD-DPPE molecule was synthesized through the conjugation of β-CD-NH

Topics: Animals; Antineoplastic Agents; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; Cell Survival; Doxorubicin; Hep G2 Cells; Humans; Hydrogen-Ion Concentration; Liver Neoplasms; MCF-7 Cells; Mice; Micelles; Nanoparticles; Organophosphorus Compounds; Tumor Microenvironment; Xenograft Model Antitumor Assays

Multifunctional nanoconjugates possessing an assortment of key functionalities such as magnetism, florescence, cell-targeting, pH and thermo-responsive features were developed for dual drug delivery. The novelty lies in careful conjugation of each of the functionality with magnetic Fe

Topics: Animals; Antineoplastic Agents; beta-Cyclodextrins; Carcinoma, Hepatocellular; Curcumin; Doxorubicin; Drug Carriers; Drug Liberation; Female; Fluoresceins; Fluorescence; Fluorescent Dyes; HeLa Cells; Humans; Magnetite Nanoparticles; Male; Mice, Inbred BALB C; Nanoconjugates; Theranostic Nanomedicine

Doxorubicin (DOX) is one of the most effective treatments for hepatocellular carcinoma (HCC), but is restricted by its poor pharmacokinetics. Herein, we exploited efficient targeted drug delivery systems and they have been found to be a worthy strategy for liver cancer therapy.. We investigated polymeric nanoparticles which were synthesized based on host-guest interaction between β-cyclodextrin and benzimidazole. The properties of nanoparticles with regard to size/shape, encapsulation efficiency, and drug release were investigated using conventional experiments. Cell proliferation assay in vitro, cell uptake assay, and cell apoptosis analysis were used to investigate cytotoxicity, uptake, and mechanism of targeted supramolecular prodrug complexes (TSPCs)-based self-assemblies and supramolecular prodrug complexes (SPCs)-based self-assemblies.. The pH-sensitive lactobionic acid (LA)-modified pH-sensitive self-assemblies were synthesized successfully. The results of in vitro released assay showed that the accelerated released of DOX from TSPCs-based self-assemblies with the decrease of pH value. When TSPCs-based self-assemblies were taken up by HepG2 cells, they demonstrated a faster release rate under acidic conditions and proved to have higher cytotoxicity than in the presence of LA. A mechanistic study revealed that TSPCs-based self-assemblies inhibited liver cell proliferation by inducing cell apoptosis.. The pH-sensitive nanocomplex, as liver-targeted nanoparticles, facilitated the efficacy of DOX in HepG2 cells, offering an appealing strategy for the treatment of HCC.

Topics: Antibiotics, Antineoplastic; Apoptosis; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Proliferation; Doxorubicin; Drug Delivery Systems; Drug Liberation; Humans; Hydrogen-Ion Concentration; Liver Neoplasms; Nanoparticles; Polymers; Tumor Cells, Cultured

A novel supramolecular nanoparticle system with core-shell structure was designed based on β-cyclodextrin-conjugated poly-l-lysine (PLCD) and hyaluronic acid for co-delivery of gene and chemotherapy agent targeting hepatocellular carcinoma (HCC). PLCD was synthesized by the conjugation of monoaldehyde activated β-cyclodextrin with poly-l-lysine via Shiff's base reaction. Doxorubicin, as a model therapeutic drug, was included into the hydrophobic cavity of β-cyclodextrin in PLCD through host-guest interaction. OligoRNA, as a model gene, was further condensed into the inclusion complexes by electrostatic interaction to form oligoRNA and doxorubicin co-loaded supramolecular nanoparticle system. Hyaluronic acid, which is often over-expressed by HCC cells, was coated on the surface of the above nanoparticles to construct HCC-targeted nanoparticle system. These nanoparticles had regular spherical shape with classic "core-shell" structure, and their size and zeta potential were 195.8nm and -22.7mV, respectively. The nanoparticles could effectively deliver doxorubicin and oligoRNA into HCC cells via CD44 receptor-mediated endocytosis and significantly inhibit the cell proliferation. In the nude mice bearing MHCC-97H tumor, the nanoparticles could be efficiently accumulated in the tumor, suggesting their strong hepatoma-targeting capability. These findings demonstrated that this novel supramolecular nanoparticle system had a promising potential for combining gene therapy and chemotherapy to treat HCC.

Topics: Animals; Antineoplastic Agents; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line, Tumor; Doxorubicin; Drug Compounding; Drug Delivery Systems; Drug Liberation; Drug Synergism; Gene Expression; Gene Transfer Techniques; Hep G2 Cells; Humans; Hyaluronan Receptors; Hyaluronic Acid; Kinetics; Liver Neoplasms; Mice; Mice, Nude; Nanoparticles; Oligoribonucleotides; Polylysine

Hepatocellular carcinoma (HCC) is one of the most lethal and prevalent cancers worldwide. CD147 (EMMPRIN or basigin) is a leading gene relating to hepatocarcinogenesis and metastasis, and is detected in transmembrane, exosome or circulating forms in HCC patients. The endosome recycling of CD147 further enhances the function of this oncoprotein from a dynamic perspective. However, previous studies about CD147 mainly focused on one separate form, and little attention has been paid to how the different forms of tumor-derived CD147 changes. Moreover, uncovering the roles of the residual C-terminal portion of CD147 after shedding is inevitable to fully understand CD147 promoting tumor progression. In this study, we discovered that under low-cholesterol condition, CD147 endocytosis is inhibited but its shedding mediated by ADAM10 is enhanced. Further procession of residual CD147 in the lysosome produces nuclear-localized CD147-ICD (intracellular domain of CD147), which contributes to autophagy through NF-κB-TRAIL-caspase8-ATG3 axis. As autophagy endows cancer cells with increased adaptability to chemotherapy, and HAb 18 (a specific antibody targeting CD147) inhibits CD147 shedding and sequential CD147-ICD enhances autophagy, we found the combination of HAb 18 and cisplatin exhibited marked antitumor efficiency.

Topics: ADAM10 Protein; ADP-Ribosylation Factor 6; ADP-Ribosylation Factors; Amyloid Precursor Protein Secretases; Animals; Antibodies, Monoclonal; Antineoplastic Agents; Autophagy; Basigin; Beclin-1; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line, Tumor; Cisplatin; Drug Resistance, Neoplasm; Female; Humans; Leupeptins; Liver Neoplasms; Membrane Proteins; Mice; Mice, Nude; Proteolysis; Simvastatin

Angelica sinensis (AS) is a traditional Chinese medicinal herb. Its ethanolic extract contains active ingredients, such as ferulic acid, ligustilide, and butylidenephthalide, which are hydrophobic and have inhibitory effects on hepatoma cells. To increase the aqueous solubility/dispersibility of AS extract and study the consequent inhibitory effect on hepatoma cells, the ethanolic extract of AS was complexed with 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), a cyclic oligosaccharide that has a hydrophilic outer surface and a hydrophobic central cavity. The AS-HP-β-CD complex (weight ratio of AS extract: HP-β-CD=1:5) was prepared and characterized. The effect of complexing the AS extract with HP-β-CD on Hep3B cell growth was investigated by analyzing cytotoxicity. Our results showed that cytotoxicity inhibition of AS-HP-β-CD complex was up to 94% and higher than that of AS extract (about 68%). These observations suggested that the use of HP-β-CD to stabilize AS extract in aqueous solution was possible for herbal medicine application.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Angelica sinensis; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line, Tumor; Dose-Response Relationship, Drug; Drug Combinations; Drug Synergism; Drugs, Chinese Herbal; Humans; Liver Neoplasms

Mixed copolymer nanoparticles (NPs) self-assembled from β-cyclodextrin-grafted hyperbranched polyglycerol (HPG-g-CD) and lactobionic acid (LA)-grafted hyperbranched polyglycerol (HPG-g-LA) were applied as carriers for a hydrophobic antitumor drug, paclitaxel (PTX), achieving hepatocellular carcinoma-targeted delivery. The resulting NPs exhibited high drug loading capacity and substantial stability in aqueous solution. In vitro drug release studies demonstrated a controlled drug release profile with increased release at acidic pH. Remarkably, tumor proliferation assays showed that PTX-loaded mixed copolymer NPs inhibited asialoglycoprotein (ASGP) receptor positive HepG2 cell proliferation in a concentration-dependent manner in comparison with ASGP receptor negative BGC-823 cells. Moreover, the competition assay demonstrated that the small molecular LA inhibited the cellular uptake of the PTX-loaded mixed copolymer NPs, indicating the ASGP receptor-mediated endocytosis in HepG2 cells. In addition, the intracellular uptake tests by confocal laser scanning microscopy showed that the mixed copolymer NPs were more efficiently taken up by HepG2 cells compared with HPG-g-CD NPs. These results suggest a feasible application of the mixed copolymer NPs as nanocarriers for hepatoma-targeted delivery of potent antitumor drugs.

Topics: Antineoplastic Agents, Phytogenic; Asialoglycoprotein Receptor; beta-Cyclodextrins; Binding, Competitive; Carcinoma, Hepatocellular; Cell Proliferation; Cell Survival; Chemistry, Pharmaceutical; Delayed-Action Preparations; Disaccharides; Dose-Response Relationship, Drug; Drug Carriers; Drug Stability; Endocytosis; Feasibility Studies; Glycerol; Hep G2 Cells; Humans; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Liver Neoplasms; Micelles; Microscopy, Confocal; Molecular Structure; Nanoparticles; Paclitaxel; Polymers; Solubility; Technology, Pharmaceutical; Time Factors

The use of active components from important medical herbs has proved effective in treating various cancers. Glycyrrhizic acid (GA) is one of the many interesting triterpenoic acids with anticancerogenic potential, and is known to trigger apoptosis in hepatocarcinoma cells. In this study we combined quantum dots (QDs) with GA in the presence of β-cyclodextrin (β-CD), and prepared β-CD/GA-functionalised QDs, which led to improved antitumor activity and induced apoptosis in hepatocarcinoma cells. These compounds showed a better selectivity for hepatic cells compared to HeLa and ECV-304 cells. Hoechst and annexin V-FITC staining and mitochondrial membrane potential (MMP) experiments proved an apoptotic effect of these compounds on HepG2 cells. At the same time, transmission electron microscopy (TEM) showed obvious features of apoptosis, for example, irregularities of nuclear shapes, mitochondria swelling, clumping and peripheral chromatin condensation, zeiosis or blebbing of the plasma membrane and formation of apoptotic bodies. It is notable that β-CD/GA-functionalised QDs showed effective cell growth inhibition by triggering G0/G1 phase arrest and inducing apoptosis through an reactive oxygen species mediated mitochondrial dysfunction pathway. β-CD/GA-functionalised QDs primarily induced apoptotic response in a time- and dose-dependent manner, but little apoptosis appeared with L-Cys-β-CD-functionalised QDs or GA alone. These studies suggest that β-CD/GA-functionalised QDs have therapeutic potential against cancer.

Topics: Apoptosis; beta-Cyclodextrins; Carcinoma, Hepatocellular; Dose-Response Relationship, Drug; G1 Phase; Glycyrrhizic Acid; HeLa Cells; Hep G2 Cells; Humans; Liver Neoplasms; Mitochondria; Quantum Dots; Reactive Oxygen Species; Resting Phase, Cell Cycle

In this study, we establish that cholesterol and sphingolipid associated with hepatitis C virus (HCV) particles are important for virion maturation and infectivity. In a recently developed culture system enabling study of the complete life cycle of HCV, mature virions were enriched with cholesterol as assessed by the molar ratio of cholesterol to phospholipid in virion and cell membranes. Depletion of cholesterol from the virus or hydrolysis of virion-associated sphingomyelin almost completely abolished HCV infectivity. Supplementation of cholesterol-depleted virus with exogenous cholesterol enhanced infectivity to a level equivalent to that of the untreated control. Cholesterol-depleted or sphingomyelin-hydrolyzed virus had markedly defective internalization, but no influence on cell attachment was observed. Significant portions of HCV structural proteins partitioned into cellular detergent-resistant, lipid-raft-like membranes. Combined with the observation that inhibitors of the sphingolipid biosynthetic pathway block virion production, but not RNA accumulation, in a JFH-1 isolate, our findings suggest that alteration of the lipid composition of HCV particles might be a useful approach in the design of anti-HCV therapy.

Topics: beta-Cyclodextrins; Carcinoma, Hepatocellular; Cell Line; Cell Line, Tumor; Cholesterol; Hepatitis C; Humans; Kidney; Liver Neoplasms; Membrane Microdomains; Plasmids; Sphingolipids; Sphingomyelin Phosphodiesterase; Virion

The mechanism(s) of fatty acid uptake by liver cells is not fully understood. We applied new approaches to address long-standing controversies of fatty acid uptake and to distinguish diffusion and protein-based mechanisms. Using HepG2 cells containing an entrapped pH-sensing fluorescence dye, we showed that the addition of oleate (unbound or bound to cyclodextrin) to the external buffer caused a rapid (seconds) and dose-dependent decrease in intracellular pH (pH(in)), indicating diffusion of fatty acids across the plasma membrane. pH(in) returned to its initial value with a time course (in min) that paralleled the metabolism of radiolabeled oleate. Preincubation of cells with the inhibitors phloretin or triacsin C had no effect on the rapid pH(in) drop after the addition of oleate but greatly suppressed pH(in) recovery. Using radiolabeled oleate, we showed that its esterification was almost completely inhibited by phloretin or triacsin C, supporting the correlation between pH(in) recovery and metabolism. We then used a dual-fluorescence assay to study the interaction between HepG2 cells and cis-parinaric acid (PA), a naturally fluorescent but slowly metabolized fatty acid. The fluorescence of PA increased rapidly upon its addition to cells, indicating rapid binding to the plasma membrane; pH(in) decreased rapidly and simultaneously but did not recover within 5 min. Phloretin had no effect on the PA-mediated pH(in) drop or its slow recovery but decreased the absolute fluorescence of membrane-bound PA. Our results show that natural fatty acids rapidly bind to, and diffuse through, the plasma membrane without hindrance by metabolic inhibitors or by an inhibitor of putative membrane-bound fatty acid transporters.

Topics: beta-Cyclodextrins; Biological Transport, Active; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Membrane; Coenzyme A Ligases; Diffusion; Fatty Acids, Unsaturated; Fluoresceins; Fluorescent Dyes; Humans; Hydrogen-Ion Concentration; Oleic Acid; Phloretin; Triazenes

The hepatitis C virus (HCV) replicates on a membrane protein complex composed of viral proteins, replicating RNA, and altered cellular membranes. Small-molecule inhibitors of cellular lipid-cholesterol metabolism such as 25-hydroxycholesterol, cerulenin, lovastatin, and GGTI-286 all show a negative effect on HCV replication. Perturbation of host cell lipid and cholesterol metabolism can disrupt replication complexes by altering membranous structures where replication occurs. Changes in cholesterol and (or) lipid composition can have a general effect on membrane structure. Alternatively, metabolic changes can exert a more subtle influence over replication complexes by altering localization of host proteins through alterations in lipid anchoring. Here, we use Huh-7 cells harboring subgenomic HCV replicons to demonstrate that 25-hydroxycholesterol, cerulenin, lovastatin, and GGTI-286 do not disrupt the membranous web where replication occurs, whereas cholesterol-depleting agents such as beta-cyclodextrin do. Cellular imaging suggests that the HCV RNA can remain associated with subcellular compartments connected with replication complexes in the presence of metabolic inhibitors. Therefore, at least 2 different molecular mechanisms are possible for the inhibition of HCV replication through the modulation of cellular lipid and cholesterol metabolism.

Topics: Anticholesteremic Agents; beta-Cyclodextrins; Carcinoma, Hepatocellular; Cells, Cultured; Cellular Structures; Cholesterol; Genome, Viral; Hepacivirus; Host-Parasite Interactions; Humans; Hydroxycholesterols; Lipid Metabolism; Lipids; Replicon; RNA, Viral; Virus Replication

Previous studies from this laboratory have demonstrated that exposure of tissue culture cells to cyclodextrins results in rapid cholesterol depletion. In the present study, we have developed experimental systems for using solutions of cyclodextrins, either 2-hydroxypropyl beta-cyclodextrin or methylated beta-cyclodextrin, complexed with varying amounts of free cholesterol to manipulate cell cholesterol content. Cholesterol delivered via the cyclodextrin has been found to be metabolically active, as measured by the acyl-coenzyme A:cholesterol acyltransferase (ACAT)-mediated esterification of [3H]cholesterol in Fu5AH rat hepatoma cells and Chinese hamster ovary cells. The methylated beta-cyclodextrin was found to be a more efficient donor in all cell types studied, with an average cholesterol uptake of at least 100 microg cholesterol/mg protein within 6 h. By modifying the cyclodextrin:cholesterol molar ratio, it is possible to manipulate the cellular cholesterol content of cells, producing conditions ranging from net cholesterol enrichment to depletion. The use of cyclodextrins provides a convenient, precise and reproducible method for modulating the cholesterol content of tissue culture cells.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Animals; beta-Cyclodextrins; Carcinoma, Hepatocellular; CHO Cells; Cholesterol; Cricetinae; Cyclodextrins; Rats; Solubility; Sterol O-Acyltransferase; Tumor Cells, Cultured