betadex and Breast-Neoplasms

In the search for photosensitizers with chemical handles to facilitate their integration into complex drug delivery nanosystems, new, unsymmetrically substituted, water insoluble meso-tetraphenylporphyrin and meso-tetra(m-hydroxyphenyl)porphyrin derivatives bearing one carboxyalkyl side chain were synthesized. Permethyl-β-cyclodextrin (pMβCD) was their ideal monomerizing host and highly efficient shuttle to transfer them into water. New assembly modes of the extremely stable (K

Topics: beta-Cyclodextrins; Biological Transport; Breast Neoplasms; Drug Carriers; Drug Delivery Systems; Endoplasmic Reticulum; Female; Humans; Magnetic Resonance Spectroscopy; MCF-7 Cells; Photosensitizing Agents; Porphyrins; Solubility; Spectrometry, Fluorescence; Water

The purpose of this study was to evaluate the potential of a newly modified cyclodextrin derivative, water-soluble β-cyclodextrin-epichlorohydrin (β-CD), as an effective drug carrier to enhance the poor solubility and bioavailability of galangin (GAL), a poorly water-soluble model drug. In this regard, inclusion complexes of GAL/β-CDP were prepared. UV-VIS spectrophotometry, Fourier-transform infrared spectroscopy (FTIR), X-ray crystallography (XRD), zeta potential analysis, particle size analysis, field emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM) were applied to characterize the synthesized GAL/β-CD. Michigan Cancer Foundation-7 (MCF-7; human breast cancer cells) and rat embryo fibroblast (REF; normal cells) were employed to examine the in vitro cytotoxic effects of GAL/β-CD using various parameters. The dye-based tests of MTT and crystal violet clearly exhibited that GAL/β-CD-treated cells had a reduced proliferation rate, an influence that was not found in the normal cell line. The cells' death was found to follow apoptotic mechanisms, as revealed by the dye-based test of acridine orange/ethidium bromide (AO/EtBr), with the involvement of the mitochondria via caspase-3-mediated events, as manifested by the Rh 123 test. We also included a mouse model to examine possible in vivo toxic effects of GAL/β-CD. It appears that the inclusion complex does not have a significant influence on normal cells, as indicated by serum levels of kidney and liver enzymatic markers, as well as thymic and splenic mass indices. A similar conclusion was reached on the histological level, as manifested by the absence of pathological alterations in the liver, kidney, thymus, spleen, heart, and lung.

Topics: Animals; beta-Cyclodextrins; Breast Neoplasms; Calorimetry, Differential Scanning; Drug Carriers; Female; Flavonoids; Humans; Mice; Rats; Solubility; Spectroscopy, Fourier Transform Infrared; Water; X-Ray Diffraction

The present investigation demonstrates the development of crosslinked β-cyclodextrin nanoparticles (β-CD NPs) for enhancing the therapeutic efficacy of docetaxel (DTX) against breast cancer. Initially, a partial inclusion complex between β-CD and polypropylene glycol (PPG) was formed to induce self-assembly. This was followed by crosslinking of β-CDs using epichlorohydrin (EPI) and removal (by solubilization) of PPG to yield uniform β-CD NPs. The formed particles were used for loading DTX to form DTX β-CD NPs. The resultant DTX β-CD NPs exhibited particle size of 223.36 ± 17.73 nm with polydispersity index (PDI) of 0.13 ± 0.09 and showed entrapment efficiency of 54.53 ± 2%. Increased cell uptake (~5-fold), cytotoxicity (~3.3-fold), and apoptosis were observed in MDA-MB-231 cells when treated with DTX β-CD NPs in comparison to free DTX. Moreover, pharmacokinetic evaluation of DTX β-CD NPs revealed ~2 and ~5-fold increase in AUC

Topics: Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Docetaxel; Drug Carriers; Female; Humans; Nanoparticles

Adjuvant-aided combination chemotherapy is one of the most effective ways of cancer treatment by overcoming the multidrug resistance (MDR) and reducing the side-effects of anticancer drugs. In this study, Conferone (Conf) was used as an adjuvant in combination with Doxorubicin (Dox) for inducing apoptosis to MDA-MB-231 cells. Herein, the novel biodegradable amphiphilic β-cyclodextrin grafted poly maleate-co-PLGA was synthesized by thiol-ene addition and ring-opening process. Micelles obtained from the novel copolymer showed exceptional properties such as small size of around 34.5 nm, CMC of 0.1 μg/mL, and cell internalization of around 100% at 30 min. These novel engineered micelles were used for combination delivery of doxorubicin-conferone with high encapsulation efficiency of near 100% for both drugs. Our results show that combination delivery of Dox and Conf to MDA-MB-231 cells had synergistic effects (CI < 1). According to cell cycle and Annexin-V apoptosis analysis, Dox-Conf loaded micelle significantly induce tumor cell apoptosis (more than 98% of cells population showed apoptosis at IC

Topics: Antibiotics, Antineoplastic; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Cycle; Cell Proliferation; Coumarins; Doxorubicin; Drug Delivery Systems; Female; Humans; Hydrogen-Ion Concentration; Micelles; Tumor Cells, Cultured

To enhance drug utilization and reduce their side effects, the strategy of "tumor-triggered targeting" was introduced to fabricate dual-pH-sensitive chitosan (CHI)/mesoporous silica nanoparticle (MSN)-based anticancer drug delivery system (DDS) in this work. Model drug doxorubicin hydrochloride (DOX) was loaded in MSN, which was modified with benzimidazole (Bz) group. Then chitosan-graft-β-cyclodextrin (CHI-g-CD) was applied as the "gatekeeper" to cover MSN through host-guest interaction between β-CD and Bz. After being coated with targeting peptide adamantane-glycine-arginine-glycine-aspartic acid-serine (Ad-GRGDS), methoxy poly(ethylene glycol) benzaldehyde (mPEG-CHO) was finally grafted on CHI through the pH-sensitive benzoic imine bond. Due to the dynamic protection of PEG, the obtained carriers were "stealthy" at pH 7.4, but could reveal the shielded targeting peptide and the positive charge of CHI in the weakly acidic environment achieved a "tumor-triggered targeting". Inside cancer cells, the interaction between β-CD and Bz group could be destroyed due to the lower pH, resulted in DOX release. Both in vitro and in vivo studies proved the DDS could targeting induce cancer cell apoptosis, inhibit tumor growth, and reduce the cytotoxicity of DOX against normal cells. It is expected that the system named DOX@MSN-CHI-RGD-PEG could be a potential choice for cancer therapy.

Topics: Adamantane; Animals; Antibiotics, Antineoplastic; Benzimidazoles; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chitosan; Delayed-Action Preparations; Doxorubicin; Drug Carriers; Drug Compounding; Female; Hydrogen-Ion Concentration; Mice; Mice, Inbred BALB C; Nanoparticles; Oligopeptides; Polyethylene Glycols; Porosity; Silicates; Tumor Burden

Cyclodextrins (CDs) have beneficial characteristics for drug delivery, including hydrophobic interior surfaces. Nanocarriers with

Topics: beta-Cyclodextrins; Breast Neoplasms; Cell Survival; Doxorubicin; Drug Carriers; Drug Delivery Systems; Female; Humans; MCF-7 Cells; Microscopy, Electron, Transmission; Nanoparticles; Silicon Dioxide; Silver; Spectroscopy, Fourier Transform Infrared

Topics: 5-Methylcytosine; Alkaline Phosphatase; Avidin; beta-Cyclodextrins; Biosensing Techniques; Breast; Breast Neoplasms; Dendrimers; DNA; Electrochemical Techniques; Female; Graphite; Humans; Immunoassay; Magnetite Nanoparticles; Nanocomposites; Stomach; Stomach Neoplasms

Non-spherical structures are beneficial to advance drug delivery effectiveness compared with common spherical ones, due to increased drug loading capability, improved bonding to a vascular wall, enhanced cellular uptake efficacy and prolonged circulation times. In this study, flower-like Zinc oxide-βcyclodextrin (βCD) nanostructures functionalized by 3-mercaptopropionic acid (MPA) as a non-spherical delivery system was successfully synthesized for aqueous delivery of curcumin (CUR) to enhance its targeting, bioavailability, and release profile. Terminal carboxyl functional groups were used for the conjugation of folic acid (FA) with the aim of active targeting to folate overexpressing breast cancer cells. The in vitro experimental study and mathematical modeling of CUR release revealed a sustained release with Fickian diffusion as the major release mechanism. MTT, colony formation and Annexin-V FITC/PI assays showed the superior anticancer effect of the system compared to free CUR against breast cancer cell line MDA-MB-231 by promoting the apoptotic respond with no cytotoxic effect on HEK293 normal cells. The efficacy of targeting strategy with FA moieties was demonstrated using the augmented cellular uptake of the FA-conjugated system on overexpressed folate receptor alpha (FRα) cells (MDA-MB-468 breast cancer cell line). Furthermore, loading of CUR to the delivery systems significantly lowered the MIC values (2.5 to 5-fold) against S. aureus and E. coli the infections of which are serious problems in cancer patients. According to the results of this study, the system can serve as a promising non-spherical delivery vehicle for enhancing bioavailability and targeting of hydrophobic anticancer agents in the future.

Topics: 3-Mercaptopropionic Acid; Antineoplastic Agents; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Curcumin; Drug Carriers; Escherichia coli; Escherichia coli Infections; Female; Folic Acid; HEK293 Cells; Humans; Nanostructures; Staphylococcal Infections; Staphylococcus aureus; Zinc Oxide

Cholesterol is abundant in plasma membranes and exhibits a variety of interactions throughout the membrane. Chemical potential accounts for thermodynamic consequences of molecular interactions, and quantifies the effective concentration (i.e., activity) of any substance participating in a process. We have developed, to our knowledge, the first method to measure cholesterol chemical potential in plasma membranes. This was accomplished by complexing methyl-β-cyclodextrin with cholesterol in an aqueous solution and equilibrating it with an organic solvent containing dissolved cholesterol. The chemical potential of cholesterol was thereby equalized in the two phases. Because cholesterol is dilute in the organic phase, here activity and concentration were equivalent. This equivalence allowed the amount of cholesterol bound to methyl-β-cyclodextrin to be converted to cholesterol chemical potential. Our method was used to determine the chemical potential of cholesterol in erythrocytes and in plasma membranes of nucleated cells in culture. For erythrocytes, the chemical potential did not vary when the concentration was below a critical value. Above this value, the chemical potential progressively increased with concentration. We used standard cancer lines to characterize cholesterol chemical potential in plasma membranes of nucleated cells. This chemical potential was significantly greater for highly metastatic breast cancer cells than for nonmetastatic breast cancer cells. Chemical potential depended on density of the cancer cells. A method to alter and fix the cholesterol chemical potential to any value (i.e., a cholesterol chemical potential clamp) was also developed. Cholesterol content did not change when cells were clamped for 24-48 h. It was found that the level of activation of the transcription factor STAT3 increased with increasing cholesterol chemical potential. The cholesterol chemical potential may regulate signaling pathways.

Topics: beta-Cyclodextrins; Breast Neoplasms; Cell Membrane; Cholesterol; Erythrocytes; Female; Humans; Neoplasm Metastasis; Signal Transduction; Thermodynamics; Tumor Cells, Cultured

Codelivery of chemo-sensitizers with chemotherapeutics using combo nanomedicine is a promising platform for overcoming chemoresistance in breast cancer. However, tumor accumulation of nano-carriers based on enhanced permeability and retention (EPR) effect is confounded by heterogeneity in tumor microenvironment. Adsorption of protein corona on surface of nanoparticle boost up clearance by reticulo-endothelial system. In this study, a surface functionalized magnetic nanocomposite (NC) for codelivery of doxorubicin (DOX) and curcumin (CUR) is developed. NCs were coated with hydroxyapatite and were also cross linked with β-cyclodextrin. NCs efficiently encapsulated DOX and CUR. Release of CUR and DOX were in a sustained pH-depended pattern. β-cyclodextrin functionalization reduced protein corona according sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. As shown by flowcytometric and confocal microscopy analyses, NCs internalized efficiently by human breast carcinoma cells MCF-7 and adriamycin resistant MCF-7 (MCF-7/adr) cells. 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) test demonstrated superior cytotoxicity of DOX-CUR loaded NCs. Anti-tumor efficacy analyses confirmed reduction in relative tumor volume size (RTV%) compared to control group. Western blot analyses demonstrated marginal CUR mediated P-glycoprotein (P-gp) down regulation. DOX-CUR loaded NCs efficiently accumulated into the tumor via external magnet guidance. Nevertheless, the increased tumor accumulation did not correlate with pharmacologic responses such as RTV% and significant superiority over free DOX was not observed.

Topics: Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; beta-Cyclodextrins; Biocompatible Materials; Biological Transport; Breast Neoplasms; Curcumin; Doxorubicin; Drug Carriers; Drug Resistance, Neoplasm; Female; Gene Expression Regulation, Neoplastic; Humans; Hydrogen-Ion Concentration; Magnets; MCF-7 Cells; Mice; Mice, Inbred BALB C; Nanomedicine; Nanoparticles

Nanomedicines have achieved several breakthroughs in cancer treatment over the past decades; however, their potential immunotoxicities are ignored, which results in serious adverse effects and greatly reduces the potential in clinical translation. Herein, we innovatively develop a theranostic supramolecular polymer using β-cyclodextrin as the host and camptothecin (CPT) as the guest linked by a glutathione-cleavable disulfide bond. The supramolecular polymerization remarkably increases the solubility of CPT by a factor of 232 and effectively inhibits its lactone ring opening in physiological environment, which is favorable for intravenous formulation and maintenance of the therapeutic efficacy. Supramolecular nanoparticles can be prepared through orthogonal self-assembly driven by π-π stacking interaction, host-guest complexation, and hydrogen bonds. The sophisticated nanomedicine constructed from the obtained supramolecular polymer can be specifically delivered to tumor sites and rapidly excreted from body after drug release, thus effectively avoiding systemic toxicity, especially long-term immunotoxicity. In vivo investigations demonstrate this supramolecular nanomedicine possesses superior antitumor performance and antimetastasis capability. This pioneering example integrating the advantages of the dynamic nature of supramolecular chemistry and nanotechnology provides a promising platform for cancer theranostics.

Topics: Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Camptothecin; Female; Glutathione; HeLa Cells; Humans; Nanomedicine; Neoplasms; Positron-Emission Tomography; Solubility

Cyclodextrins (CD) are natural macrocyclic oligosaccharides linked by α(1,4) glycosidic bonds. Hydrophobic cavity of CDs are able to incorporate small molecules, ions, macromolecules which makes them excellent delegates for forming nanoparticulate carriers upon chemical modification to render amphiphilicity to CDs. In this study, blank 6OCaproβCD nanoparticle was prepared and administered to MCF-7 breast cancer cells. The effects of these nanoparticles on the cells were investigated in depth through biochemical and proteomic tests following 48 h of incubation. Proteomics studies revealed that apoptosis-related protein levels of hnRNP and CBX1 were increased while HDGF was not affected supporting the idea that 6OCaproβCD nanoparticles prevent cell proliferation. Gene expression studies were generally in correlation with protein levels since gene expression was significantly stimulated while protein levels were lower compared to the control group suggesting that a post-transcriptional modification must have occurred. Furthermore, 6OCaproβCD was observed to not trigger multidrug resistance as proved with RT-PCR that effectuates another exquisite characteristic of 6OCaproβCD nanoparticle as carrier of chemotherapeutic drugs. Metabolomic pathways of CD effect on MCF7 cells were elucidated with HMDB as serine biosynthesis, transmembrane transport of small molecules, metabolism of steroid hormones, estrogen biosynthesis and phospholipid biosynthesis. In conclusion, 6OCaproβCD is a promising nanoparticulate carrier for chemotherapeutic drugs with intrinsic apoptotic effect to be employed in treatment of breast cancer and further studies should be conducted in order to comprehend the exact mechanism of action.

Topics: Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Membrane; Cell Proliferation; Cholesterol; Chromobox Protein Homolog 5; Chromosomal Proteins, Non-Histone; Drug Carriers; Drug Compounding; Energy Metabolism; Female; Gene Expression Regulation, Neoplastic; Genomics; Heterogeneous-Nuclear Ribonucleoproteins; Humans; MCF-7 Cells; Metabolomics; Nanoparticles; Proteomics; Signal Transduction

Overexpression of RhoC protein in breast cancer patients has been linked to increased cancer cell invasion, migration, and metastases. Suppressing RhoC expression in aggressive breast cancer cells using silencing RNA (siRNA) molecules is a viable strategy to inhibit the metastatic spread of breast cancer. In this report, we describe the synthesis of a series of asymmetric pH-sensitive, membrane-destabilizing polymers engineered to complex anti-RhoC siRNA molecules forming "smart" nanoparticles. Using β-CD as the particle core, polyethylene glycol (PEG) chains were conjugated to the primary face via non-cleavable bonds and amphiphilic polymers incorporating hydrophobic and cationic monomers were grafted to the secondary face via acid-labile linkages. We investigated the effect of PEG molecular weight (2 & 5 kDa) on transfection capacity and serum stability of the formed particles. We evaluated the efficacy of EPPT1 peptides presented on the free tips of the PEG brush to function as a targeting ligand against underglycosylated MUC1 (uMUC1) receptors overexpressed on the surface of metastatic breast cancer cells. Results show that "smart" nanoparticles successfully delivered anti-RhoC siRNA into the cytoplasm of aggressive SUM149 and MDA-MB-231 breast cancer cells, which resulted in a dose-dependent inhibition of cell migration and invasion. Further, EPPT1-targeted nanoparticles demonstrate a synergistic inhibition of cell migration and invasion imparted via RhoC knockdown and EPPT1-mediated signaling via the uMUC1 receptor.

Topics: beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Membrane Permeability; Cell Movement; Cell Proliferation; Drug Liberation; Female; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Mucin-1; Nanocapsules; Neoplasm Invasiveness; Oligopeptides; Polyethylene Glycols; Polymerization; rhoC GTP-Binding Protein; RNA Interference; RNA, Small Interfering; Transfection; Tumor Microenvironment

Previous reports have documented that cholesterol-lowering simvastatin prevented osteolytic metastasis of breast cancer in animal model in which cancer cells were placed into blood circulation. Thus, simvastatin treatment might have a preventive effect in inhibiting osteoclast activity of metastatic bone microenvironment. This study documented that both simvastatin and MBCD (cholesterol depleting drug) blocked the breast cancer-induced TRAP and MMP activity, and expressions of various osteoclastogenic genes (TRAP, Cathepsin K, and NFATc1) in pre-osteoclast RAW264.7 cells, and osteoclastogenic CSF-1 and RANKL expressions in breast cancer MCF-7 cells. Thus, these findings unravel a molecular mechanism of simvastatin-/MBCD-mediated inhibition of breast cancer-driven osteoclast activity.

Topics: Animals; Anticholesteremic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Differentiation; Female; Humans; MCF-7 Cells; Mice; Osteoclasts; Signal Transduction; Simvastatin

Paclitaxel (PTX) is one of the drugs of choice in the treatment of breast and lung cancer. However, its severe side effects, including mielosuppression, cardiotoxicity and neurotoxicity, frequently cause treatment to be discontinued. Solid lipid nanoparticles (NPs) of glyceril tripalmitate (tripalmitin) loaded with PTX (Tripalm-NPs-PTX) including modifications by the addition of hexa(ethylene glycol), β-cyclodextrin and macelignan were developed. All NPs-PTX formulations displayed excellent hemocompatibility and significantly enhanced PTX antitumor activity in human breast (MCF7, MDAMB231, SKBR3 and T47D) and lung (A549, NCI-H520 and NCI-H460) cancer cells. Tripalm-NPs-PTX decreased PTX IC

Topics: Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Line; Female; Humans; Lignans; Lung Neoplasms; MCF-7 Cells; Nanoparticles; Neoplastic Stem Cells; Paclitaxel; Polyethylene Glycols; Spheroids, Cellular; Triglycerides; Tumor Cells, Cultured

Topics: Animals; beta-Cyclodextrins; Breast Neoplasms; Cytokines; Female; Inflammasomes; Inflammation; Injections, Intraperitoneal; Liver; Liver Neoplasms; Lung Neoplasms; Macrophages, Peritoneal; Mammary Neoplasms, Experimental; Mice, Inbred C57BL; Nanoparticles; NLR Family, Pyrin Domain-Containing 3 Protein; Polyethyleneimine; Spleen

Pharmaceutical industries spend more money in developing new and efficient methods for delivering successful drugs for anticancer therapy. Electrospun nanofibers and nanoparticles loaded with drugs have versatile biomedical applications ranging from wound healing to anticancer therapy. We aimed to attempt for fabricating elastomeric poly (l-lactic acid-co-ε-caprolactone) (PLACL) with Aloe Vera (AV), magnesium oxide (MgO) nanoparticles, curcumin (CUR) and β-cyclodextrin (β-CD) composite nanofibers to control the growth of MCF-7 cells for breast cancer therapy. The study focused on the interaction of MgO nanoparticle with CUR and β-CD inhibiting the proliferation of Michigan Cancer Foundation-7 (MCF-7) breast cancer cells. FESEM micrographs of fabricated electrospun PLACL, PLACL/AV, PLACL/AV/MgO, PLACL/AV/MgO/CUR and PLACL/AV/MgO/β-CD nanofibrous scaffolds achieved bead free, random and uniform nanofibers with fiber diameter in the range of 786±286, 507±171, 334±95, 360±94 and 326±80nm respectively. Proliferation of MCF-7 cells was decreased by 65.92% in PLACL/AV/MgO/CUR with respect to PLACL/AV/MgO nanofibrous scaffolds on day 9. The obtained results proved that 1% CUR interacting with MgO nanoparticles showed higher inhibition of MCF-7 cells among all other nanofibrous scaffolds thus serving as a promising biocomposite material system for the breast cancer therapy.

Topics: Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Curcumin; Female; Humans; Manganese Compounds; MCF-7 Cells; Nanofibers; Nanoparticles; Oxides; Polyesters

The development of biocompatible vector for hydrophobic drug delivery remains a longstanding issue in cancer therapy. We design and synthesis a drug delivery system based on HPG modified β-CD (β-CD-HPG) by conjugating HPG branches onto β-CD core and its structure was confirmed by NMR, FTIR, GPC and solubility. In vitro biocompatibility tests showed that HPG modification significantly improved red blood cells morphology alteration and hemolysis cause by β-CD and β-CD-HPG displayed cell safety apparently in a wide range of 0.01-1mg/mL. An anti-cancer drug, docetaxel, was effectively encapsulated into β-CD-HPG which was confirmed by DSC analysis. This copolymer could form nanoparticles with small size (<200nm) and exhibited better DTX loading capacity and controlled release kinetics without initial burst release behavior compared with β-CD. Furthermore, antitumor assay in vitro show that β-CD-HPG/DTX effectively inhibited proliferation of human breast adenocarcinoma cells. Therefore, β-CD-HPG/DTX exhibit great potential for cancer chemotherapy.

Topics: Adenocarcinoma; beta-Cyclodextrins; Breast Neoplasms; Cell Line; Delayed-Action Preparations; Docetaxel; Drug Screening Assays, Antitumor; Erythrocytes; Female; Glycerol; Hemolysis; Humans; Polymers; Taxoids

Several targeted drug delivery systems have recently been developed to increase the bioavailability of a drug at its site of action, allowing simultaneous reduction of the total necessary drug dose as well as side effects. Here, we designed a cationic gene vector containing matrix metalloproteinase-2 (MMP2)-cleavable substrate peptides that specifically target tumor sites where MMP2 levels are high. The targeted delivery system is fabricated by linking enzyme-cleavable polyethylene glycol (PEG) derivatives to cationic β-cyclodextrin-polyethylenimine conjugates, which reduce the toxicity of polyethylenimine and condense the therapeutic cargo. In the present study, tumor suppressor microRNA miR-34a, which suppresses onset and progression of many types of cancers, was investigated for its therapeutic potential for treating breast cancer. The PEG coating markedly reduces nonspecific interaction between cationic particles and serum proteins, permitting accumulation at the target site; subsequent peptide cleavage by MMP2 facilitates miR-34a delivery into tumor cells. The nanopreparation shows excellent stability, and its internalization, tumor targeting, and antitumor efficacy in vitro and in vivo are better than those of a nanopreparation containing MMP2-uncleavable peptide.

Topics: Animals; beta-Cyclodextrins; Breast Neoplasms; Cations; Cell Line; Cell Line, Tumor; Dogs; Drug Delivery Systems; Female; Genetic Vectors; Humans; Madin Darby Canine Kidney Cells; Matrix Metalloproteinase 2; MicroRNAs; Nanoparticles; Peptide Hydrolases; Peptides; Polyethylene Glycols; Polyethyleneimine

Fisetin (FST), a potent anticancer phytoconstituent, exhibits poor aqueous solubility and hence poor bioavailability. The aim of the present study is to improve the oral bioavailability of FST by encapsulating into PLGA NPs (poly-lactide-co-glycolic acid nanoparticles) as a complex of HPβCD (hydroxyl propyl beta cyclodextrin) and to assess its anti-cancer activity against breast cancer cells. FST-HPβCD inclusion complex (FHIC) was prepared and the supramolecular complex formation was characterized by FTIR, DSC, PXRD and

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Administration, Oral; Animals; Antineoplastic Agents; Apoptosis; beta-Cyclodextrins; Biological Availability; Breast Neoplasms; Cell Line, Tumor; Female; Flavonoids; Flavonols; Humans; Lactic Acid; MCF-7 Cells; Mice; Mice, Inbred C57BL; Nanoparticles; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Reactive Oxygen Species; Solubility

Liposome (spherical vesicles) and cochleate (multilayer crystalline, spiral structure) formulations containing raloxifene have been developed having dimethyl-β-cyclodextrin (DM-β-CD) or sodium taurocholate (NaTC). Raloxifene was approved initially for the treatment of osteoporosis but it is also effective on breast tissue and endometrial cells. Raloxifene inhibits matrix metalloproteinase-2 (MMP-2) enzyme, which is known to be responsible for tumor invasion and the initiation of angiogenesis during the tumor growth. Therefore, raloxifene was selected as a model drug. A series of raloxifene-loaded liposome and cochleate formulations were prepared. In vitro release studies and in vivo tests were performed. Breast cancer cell lines (MCF-7) were also used to find the most effective formulation. Highest antitumor activity was observed, and MMP-2 enzyme was also found to be inhibited with raloxifene-loaded cochleates containing DM-β-CD. These developed formulations can be helpful for further treatment alternatives and new strategies for cancer therapy.

Topics: Animals; Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Caco-2 Cells; Cell Line, Tumor; Chemistry, Pharmaceutical; Female; Humans; Liposomes; Matrix Metalloproteinase 2; MCF-7 Cells; Raloxifene Hydrochloride; Rats; Rats, Sprague-Dawley; Taurocholic Acid

The clinical application of nanoparticular Gd(III) based contrast agents for tumor molecular MRI has been hindered by safety concerns associated with prolonged tissue retention, although they can produce strong tumor enhancement. In this study, a targeted well-defined cyclodextrin-based nanoglobular contrast agent was developed through self-assembly driven by host-guest interactions for safe and effective cancer molecular MRI. Multiple β-cyclodextrins attached POSS (polyhedral oligomeric silsesquioxane) nanoglobule was used as host molecule. Adamantane-modified macrocyclic Gd(III) contrast agent, cRGD (cyclic RGDfK peptide) targeting ligand and fluorescent probe was used as guest molecules. The targeted host-guest nanoglobular contrast agent cRGD-POSS-βCD-(DOTA-Gd) specifically bond to αvβ3 integrin in malignant 4T1 breast tumor and provided greater contrast enhancement than the corresponding non-targeted agent. The agent also provided significant fluorescence signal in tumor tissue. The histological analysis of the tumor tissue confirmed its specific and effective targeting to αvβ3 integrin. The targeted imaging agent has a potential for specific cancer molecular MR and fluorescent imaging.

Topics: Animals; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Contrast Media; Disease Models, Animal; Female; Integrin alphaVbeta3; Magnetic Resonance Imaging; Mice; Mice, Inbred BALB C; Molecular Imaging; Nanostructures; Peptides, Cyclic

Cancer metastasis is the leading reason for the high mortality of breast cancer. Herein, we report on a pH-responsive host-guest nanosystem of succinobucol (PHN) with pH-stimuli controlled drug release behavior to improve the therapeutic efficacy on lung metastasis of breast cancer. PHN was composed of the host polymer of β-cyclodextrin linked with multiple arms of N,N-diisopropylethylenediamine (βCD-DPA), the guest polymer of adamantyl end-capped methoxy poly(ethylene glycol) (mPEG-Ad), and the active agent of succinobucol. PHN comprises nanometer-sized homogenous spherical particles, and exhibits specific and rapid drug release in response to the intracellular acidic pH-stimuli. Then, the anti-metastatic efficacy of PHN is measured in metastatic 4T1 breast cancer cells, which effectively confirms the superior inhibitory effects on cell migration and invasion activities, VCAM-1 expression and cell-cell binding of RAW 264.7 to 4T1 cells. Moreover, PHN can be specifically delivered to the sites of metastatic nodules in lungs, and result in an obviously improved therapeutic efficacy on lung metastasis of breast cancer. Thereby, the pH-responsive host-guest nanosystem can be a promising drug delivery platform for effective treatment of cancer metastasis.

Topics: Animals; Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Movement; Disease Models, Animal; Drug Delivery Systems; Female; Hydrogen-Ion Concentration; Lung Neoplasms; Mice, Inbred BALB C; Nanostructures; Neoplasm Invasiveness; Polyethylene Glycols; Probucol; Treatment Outcome

The potent and selective anti-tumor agent, di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone (Dp44mT), localizes in lysosomes and forms cytotoxic copper complexes that generate reactive oxygen species (ROS), resulting in lysosomal membrane permeabilization (LMP) and cell death. Herein, the role of lysosomal membrane stability in the anti-tumor activity of Dp44mT was investigated. Studies were performed using molecules that protect lysosomal membranes against Dp44mT-induced LMP, namely heat shock protein 70 (HSP70) and cholesterol. Up-regulation or silencing of HSP70 expression did not affect Dp44mT-induced LMP in MCF7 cells. In contrast, cholesterol accumulation in lysosomes induced by the well characterized cholesterol transport inhibitor, 3-β-[2-(diethyl-amino)ethoxy]androst-5-en-17-one (U18666A), inhibited Dp44mT-induced LMP and markedly and significantly (p<0.001) reduced the ability of Dp44mT to inhibit cancer cell proliferation (i.e., increased the IC(50)) by 140-fold. On the other hand, cholesterol extraction using methyl-β-cyclodextrin enhanced Dp44mT-induced LMP and significantly (p<0.01) increased its anti-proliferative activity. The protective effect of U18666A in increasing lysosomal cholesterol and preventing the cytotoxic activity of Dp44mT was not due to induced autophagy. Instead, U18666A was found to decrease lysosomal turnover, resulting in autophagosome accumulation. Moreover, preincubation with U18666A did not prevent the ability of Dp44mT to induce autophagosome synthesis, indicating that autophagic initiation via Dp44mT occurs independently of LMP. These studies demonstrate the significance of lysosomal membrane stability in relation to the ability of Dp44mT to execute tumor cell death and overcome pro-survival autophagy. Hence, lysosomal-dependent cell death induced by Dp44mT serves as an important anti-tumor strategy. These results are important for comprehensively understanding the mechanism of action of Dp44mT.

Topics: Androstenes; Anticholesteremic Agents; Antineoplastic Agents; Autophagy; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Cholesterol; Dose-Response Relationship, Drug; Female; HSP70 Heat-Shock Proteins; Humans; Inhibitory Concentration 50; Intracellular Membranes; Lysosomes; MCF-7 Cells; Permeability; RNA Interference; Thiosemicarbazones; Transfection

A near infrared (NIR) light responsive chromophore 7-(diethylamino)-4-(hydroxymethyl)-2H-chromen-2-one (DEACM) was synthesized and incorporated to β-cyclodextrins with cRGD functionalized poly(ethylene glycol), the amphiphiles were coordinated with Au nanorods or nanoparticles to load anticancer drug doxorubicin (DOX) for fabricating hybrid nanoparticles. The π-π stacking interaction between DEACM and DOX was formed in the hybrid nanoparticles, which contributed to the high drug loading content. The Au nanorods or nanoparticles enhanced the photosolvolysis of DEACM under the irradiation of NIR with 808 nm wavelength and triggered the accelerated drug release from the nanoparticles. The drug loaded hybrid nanoparticles with NIR irradiation exhibited efficient inhibition effect on the proliferation of 4T1 breast cancer cells in vitro. The in vivo anticancer activity study on breast cancer bearing mice revealed that the hybrid nanoparticles containing Au nanorods exhibited excellent anticancer activity under the irradiation of 808 nm wavelength NIR with 800 mW.

Topics: Animals; Antineoplastic Agents; Benzopyrans; beta-Cyclodextrins; Breast Neoplasms; Breast Neoplasms, Male; Cell Line, Tumor; Cell Proliferation; Doxorubicin; Drug Delivery Systems; Female; Gold; Infrared Rays; Male; Mice; Mice, Inbred BALB C; Nanoparticles; Nanotubes; NIH 3T3 Cells; Peptides, Cyclic; Polyethylene Glycols

Recently, biologically active compounds isolated from plants used in herbal medicine have been the center of interest. Deoxypodophyllotoxin (DPT), structurally closely related to the lignan podophyllotoxin, was found to be a potent antitumor and antiproliferative agent, in several tumor cells, in vitro. However, DPT has not been used clinically yet because of the lack of in vivo studies. This study is the first report demonstrating the antitumor effect of DPT on MDA-MB-231 human breast cancer xenografts in nude mice. DPT, significantly, inhibited the growth of MDA-MB-231 xenograft in BALB/c nude mice. The T/C value (the value of the relative tumor volume of treatment group compared to the control group) of groups treated with 5, 10, and 20 mg/kg of intravenous DPT-HP-β-CD was 42.87%, 34.04% and 9.63%, respectively, suggesting the positive antitumor activity of DPT. In addition, the antitumor effect of DPT-HP-β-CD (20 mg/kg) in human breast cancer MDA-MB-231 xenograft was more effective than etoposide (VP-16) (20 mg/kg) and docetaxel (20 mg/kg). These findings suggest that this drug is a promising chemotherapy candidate against human breast carcinoma.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Administration, Intravenous; Animals; Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Docetaxel; Drugs, Chinese Herbal; Etoposide; Female; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Podophyllotoxin; Taxoids; Xenograft Model Antitumor Assays

Manipulation of cell-cell interactions has potential applications in basic research and cell-based therapy. Herein, using a combination of metabolic glycan labelling and bio-orthogonal click reaction, we engineer cell membranes with β-cyclodextrin and subsequently manipulate cell behaviours via photo-responsive host-guest recognition. With this methodology, we demonstrate reversible manipulation of cell assembly and disassembly. The method enables light-controllable reversible assembly of cell-cell adhesion, in contrast with previously reported irreversible effects, in which altered structure could not be reused. We also illustrate the utility of the method by designing a cell-based therapy. Peripheral blood mononuclear cells modified with aptamer are effectively redirected towards target cells, resulting in enhanced cell apoptosis. Our approach allows precise control of reversible cell-cell interactions and we expect that it will promote further developments of cell-based therapy.

Topics: Apoptosis; Azo Compounds; beta-Cyclodextrins; Biomedical Engineering; Breast Neoplasms; Cell Adhesion; Cell Communication; Cell Membrane; Female; Flow Cytometry; HeLa Cells; Humans; Leukocytes, Mononuclear; Ligands; MCF-7 Cells; Polymers; Polysaccharides; Spatio-Temporal Analysis

A programmed supramolecular nanocarrier was developed for multistage targeted photodynamic therapy. This smart nanocarrier exhibited enhanced cellular uptake and controlled mitochondria targeting, as well as an excellent photodynamic therapeutic effect after light irradiation.

Topics: Amino Acid Sequence; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Chlorophyllides; Drug Carriers; Female; Humans; Light; MCF-7 Cells; Microscopy, Confocal; Mitochondria; Nanoparticles; Peptides; Photochemotherapy; Photosensitizing Agents; Porphyrins

Multivalent mannose-functionalized nanoparticles self-assembled from amphiphilic β-cyclodextrins (β-CDs) facilitate the targeted delivery of anticancer drugs to specific cancer cells. Doxorubicin (DOX)-loaded nanoparticles equipped with multivalent mannose target units were efficiently taken up via receptor-mediated endocytosis by MDA-MB-231 breast cancer cells that overexpress the mannose receptor. Upon entering the cell, the intracellular pH causes the release of DOX, which triggers apoptosis. Targeting by multivalent mannose significantly improved the capability of DOX-loaded nanoparticles to inhibit the growth of MDA-MB-231 cancer cells with minimal side effects in vivo. This targeted and controlled drug delivery system holds promise as a nanotherapeutic for cancer treatment.

Topics: Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Doxorubicin; Drug Delivery Systems; Female; Humans; Lectins, C-Type; Mannose; Mannose Receptor; Mannose-Binding Lectins; Nanoparticles; Receptors, Cell Surface

The targeted delivery of a photosensitizer (PS) into specific cancer cells is an effective way to enhance the efficacy and minimize the side effects of photodynamic therapy. Herein, heptamannosylated β-cyclodextrin (β-CD) was used to mediate the formation of an adamantane (Ad)-functionalized BODIPY PS nanoparticle via strong β-CD/Ad complexation. The mannose-functionalized PS nanoparticles are selectively internalized by mannose-receptor-rich MDA-MB-231 breast cancer cells via receptor-mediated endocytosis, facilitating singlet oxygen generation to trigger apoptosis in cancer cells upon red-light irradiation. These nanoparticles exhibit excellent targeted delivery of the PS, leading to cancer cell death after irradiation both in vitro and in vivo.

Topics: Adamantane; beta-Cyclodextrins; Boron Compounds; Breast Neoplasms; Cell Death; Humans; Nanoparticles; Photochemotherapy; Photosensitizing Agents

Aptamers are single-stranded RNA or DNA ligands that can specifically bind to various molecular targets with high affinity. Owing to this unique character, they have become increasingly attractive in the field of drug delivery. In this study, we developed a multifunctional composite micelle (CM) with surface modification of aptamer AS1411 (Ap) for targeted delivery of doxorubicin (DOX) to human breast tumors. This binary mixed system consisting of AS1411 modified Pluronic F127 and beta-cyclodextrin-linked poly(ethylene glycol)-b-polylactide could enhance DOX-loading capacity and increase micelle stability. Cellular uptake of CM-Ap was found to be higher than that of untargeted CM due to the nucleolin-mediated endocytosis effect. In vivo study in MCF-7 tumor-bearing mice demonstrated that the AS1411-functionalized composite micelles showed prolonged circulation time in blood, enhanced accumulation in tumor, improved antitumor activity, and decreased cardiotoxicity. In conclusion, aptamer-conjugated multifunctional composite micelles could be a potential delivery vehicle for cancer therapy.

Topics: Animals; Antineoplastic Agents; Aptamers, Nucleotide; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cyclodextrins; Doxorubicin; Drug Delivery Systems; Female; Humans; Mice; Micelles; Nanomedicine; Neoplasm Transplantation; Oligodeoxyribonucleotides; Poloxamer; Polyethylene Glycols

We examined the phytoestrogenic effects of palmiwon on breast carcinoma, lipid accumulation in methyl-β-cyclodextrin-induced HepG2 cells, and lipid-related diseases in a rat model of menopausal hyperlipidemia.. E-Screen assay was used to screen for phytoestrogens, especially those with antiestrogenic activity, in MCF-7 cells. Oil Red O staining and intracellular cholesterol analyses were used to quantify cellular cholesterol levels. 3-Hydroxy-3-methyl glutaryl coenzyme A reductase assay was used to measure enzyme activity. The levels of phosphorylated adenosine monophosphate-activated protein kinases and products of genes involved in cholesterol synthesis were measured by Western blot analysis. Thirty rats were either ovariectomized or sham-operated and randomly assigned to four groups (n = 5)-Sham, OVX, OVX-SV, or OVX-PMW (50, 150, or 450 mg/kg) group-for 8 weeks. A number of targets associated with lipid-related diseases were examined to confirm the estrogenic effects of palmiwon.. Palmiwon showed antiestrogenic activity in MCF-7 cells. Palmiwon decreased lipid accumulation, total cholesterol levels, and low-density lipoprotein/very-low-density lipoprotein levels in HepG2 cells. Moreover, palmiwon reversed the effects of methyl-β-cyclodextrin on cholesterol synthesis regulators and inhibited the activity of 3-hydroxy-3-methyl glutaryl coenzyme A reductase. Phosphorylation of adenosine monophosphate-activated protein kinase was stimulated by palmiwon. In ovariectomized rats, palmiwon reduced retroperitoneal and perirenal fat accumulation, serum lipids, atherogenic index, cardiac risk factor score, intima-media thickness, and nonalcoholic steatohepatitis scores.. These results indicate that palmiwon inhibits lipid accumulation without estrogenic activity in the breast. Therefore, palmiwon may have potential as a therapeutic agent for the treatment of hyperlipidemia in postmenopausal women.

Topics: Animals; beta-Cyclodextrins; Breast Neoplasms; Carotid Intima-Media Thickness; Cholesterol; Cyclic AMP-Dependent Protein Kinases; Drugs, Chinese Herbal; Estrogens; Female; Hep G2 Cells; Humans; Hydroxymethylglutaryl CoA Reductases; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hyperlipidemias; Lipid Metabolism; Lipids; Lipoproteins, LDL; MCF-7 Cells; Menopause; Models, Animal; Phosphorylation; Phytoestrogens; Rats; Rats, Sprague-Dawley

Paclitaxel (PTX) solid lipid nanoparticles (SLNs) modified with 2-hydroxypropyl-β-cyclodextrin (HPCD) were evaluated for their ability to enhance PTX absorption and reduce the nephrotoxicity accompanying intravenous administration.. PTX-loaded SLNs (PS) and PTX-loaded SLNs modified using HPCD (PSC) were prepared by hot-melted sonication. The anticancer activity of PSC was evaluated in MCF-7 cells, and confocal microscopy was used to quantify the cellular uptake. The pharmacokinetic profiles of PTX released from PSC after intravenous administration were studied in rats. Furthermore, kidney toxicity was determined by measuring the kidney size and plasma creatinine level.. PSC were successfully prepared by hot-melted sonication and had smaller diameters than PS. PSC exhibited improved anticancer activity and cellular uptake in MCF-7 cells. Furthermore, PSC showed higher bioavailability in rats after intravenous administration than PTX solution; however, no significant differences in kidney toxicity were observed.. Based on these results, PSC could be considered as a potential therapeutic PTX delivery system for breast cancer with low renal toxicity.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Albumins; Animals; Antineoplastic Agents; Apoptosis; Area Under Curve; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Cell Survival; Creatinine; Drug Carriers; Drug Screening Assays, Antitumor; Female; Flow Cytometry; Humans; Injections, Intravenous; Kidney; Lipids; Male; MCF-7 Cells; Mice; Mice, Inbred BALB C; Microscopy, Confocal; Nanoparticles; Neoplasm Transplantation; Paclitaxel; Rats

Studies have shown that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in cancer cells. However, breast cancer cells are generally resistant to TRAIL. In the present study, we explored the effect of bufalin on TRAIL-induced breast cancer cell apoptosis. The results showed that bufalin enhanced TRAIL-induced apoptosis in MCF-7 and MDA-MB-231 breast cancer cells by activating the extrinsic apoptotic pathway. Bufalin also promoted the clustering of death receptor 4 (DR4) and DR5 in aggregated lipid rafts. The cholesterol-sequestering agent methyl-β-cyclodextrin reversed the DR4 and DR5 clustering and reduced bufalin+TRAIL-induced apoptosis. Overall, these results indicate that bufalin enhanced TRAIL-induced apoptosis in breast cancer cells by the partial redistribution of DRs in lipid rafts.

Topics: Antineoplastic Agents; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Bufanolides; Cell Line, Tumor; Cholesterol; Female; Humans; Membrane Microdomains; Receptors, TNF-Related Apoptosis-Inducing Ligand; TNF-Related Apoptosis-Inducing Ligand

The vasculature in tumor microenvironment plays important roles in the tumor growth and metastasis, and the combination of vascular disrupting agents with chemotherapeutic drugs should be effective in inhibiting tumor progression. But the dosing schedules are essential to achieve a balance between vascular collapse and intratumoral uptake of chemotherapeutic agents. In the current study, emulsion and blend electrospinning were used to create compartmental fibers accommodating both combretastatin A-4 (CA4) and hydroxycamptothecin (HCPT). The release durations of CA4 and HCPT were modulated through the structure of fibers for dual drug loadings and the inoculation of 2-hydroxypropyl-β-cyclodextrin in fiber matrices. Under a noncontact cell coculture in Transwell, the sequential release of CA4 and HCPT indicated a sequential killing of endothelial and tumor cells. In an orthotopic breast tumor model, all the CA4/HCPT-loaded fibers showed superior antitumor efficacy and higher survival rate than fibers with loaded individual drug. Compared with fibrous mats with infiltrated free CA4 and fibers with extended release of CA4 for over 30 days, fibers with sustained release of CA4 for 3-7 days from CA4/HCPT-loaded fibers resulted in the most significant antitumor efficacy, tumor vasculature destruction, and the least tumor metastasis to lungs. A judicious selection of CA4 release durations in the combination therapy should be essential to enhance the tumor suppression efficacy and antimetastasis activity.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Camptothecin; Drug Carriers; Drug Liberation; Female; Lactates; Lung Neoplasms; Mice; Mice, Inbred BALB C; Polyethylene Glycols; Stilbenes

HSP90 may be overexpressed in cancer cells which are greatly dependent on Hsp90 function. Geldanamycin derivative 17 allylamino-17-demethoxygeldanamycin (17-AAG) inhibits the function and expression of HSP90. 17-AAG has poor water-solubility which is a potential problem for clinical practice. In this study for improving the stability and solubility of molecules in drug delivery systems we used a β-cyclodextrin- 17AAG complex.. To assess cytotoxic effects of β-cyclodextrin-17AAG complexes and free 17AAG, colorimetric cell viability (MTT) assays were performed. Cells were treated with equal concentrations of β-cyclodextrin- 17AAG complex and free 17AAG and Hsp90 gene expression levels in the two groups was compared by real-time PCR.. MTT assay confirmed that β-cyclodextrin- 17AAG complex enhanced 17AAG cytotoxicity and drug delivery in T47D breast cancer cells. The level of Hsp90 gene expression in cells treated with β-cyclodextrin- 17AAG complex was lower than that of cells treated with free 17AAG (P=0.001).. The results demonstrated that β-cyclodextrin- 17AAG complexes are more effective than free 17AAG in down-regulating HSP90 expression due to enhanced β-cyclodextrin-17AAG uptake by cells. Therefore, β-cyclodextrin could be superior carrier for this kind of hydrophobic agent.

Topics: Benzoquinones; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Drug Screening Assays, Antitumor; Female; Gene Expression Regulation, Neoplastic; HSP90 Heat-Shock Proteins; Humans; Lactams, Macrocyclic; Nanoparticles; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Nowadays, the encapsulation of cytotoxic chemotherapeutic agents is attracting interest as a method for drug delivery. We hypothesized that the efficiency of helenalin might be maximized by encapsulation in β-cyclodextrin nanoparticles. Helenalin, with a hydrophobic structure obtained from flowers of Arnica chamissonis and Arnica Montana, has anti-cancer and anti-inflammatory activity but low water solubility and bioavailability. β-Cyclodextrin (β-CD) is a cyclic oligosaccharide comprising seven D-glucopyranoside units, linked through 1,4-glycosidic bonds.. To test our hypothesis, we prepared β-cyclodextrin- helenalin complexes to determine their inhibitory effects on telomerase gene expression by real-time polymerase chain reaction (q-PCR) and cytotoxic effects by colorimetric cell viability (MTT) assay.. MTT assay showed that not only β-cyclodextrin has no cytotoxic effect on its own but also it demonstrated that β-cyclodextrin- helenalin complexes inhibited the growth of the T47D breast cancer cell line in a time and dose-dependent manner. Our q-PCR results showed that the expression of telomerase gene was effectively reduced as the concentration of β-cyclodextrin-helenalin complexes increased.. β-Cyclodextrin-helenalin complexes exerted cytotoxic effects on T47D cells through down-regulation of telomerase expression and by enhancing Helenalin uptake by cells. Therefore, β-cyclodextrin could be superior carrier for this kind of hydrophobic agent.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Female; Humans; Nanoparticles; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sequestering Agents; Sesquiterpenes; Sesquiterpenes, Guaiane; Spectroscopy, Fourier Transform Infrared; Telomerase; Tumor Cells, Cultured

This study aimed to evaluate the potential of solid lipid nanoparticles (SLNs) of paclitaxel (PTX) modified with a 2-hydroxypropyl-β-cyclodextrin system to enhance cellular accumulation of PTX into p-glycoprotein (p-gp)-expressing cells.. The PTX-loaded-SLNs consisted of lipid (stearic acid) and surfactants (lecithin and poloxamer 188) and were then modified with 2-hydroxypropyl-β-cyclodextrin by a sonication method.. In terms of cytotoxicity, PTX-loaded SLNs modified with 2-hydroxypropyl-β-cyclodextrin showed higher cytotoxicity than other formulations. In particular, the cellular uptake of PTX from PTX-loaded SLNs modified with 2-hydroxypropyl-β-cyclodextrin was about 5.8- and 1.5-fold higher than that from PTX solution and unmodified PTX-loaded SLNs in MCF-7/ADR cells, respectively. After a 4-h incubation, clear fluorescence images inside cells were observed over time. When PTX-loaded SLNs modified with 2-hydroxypropyl-β-cyclodextrin were incubated with MCF-7/ADR cells for 4 h, cellular uptake of PTX increased 1.7-fold versus that of PTX in the presence of verapamil.. These results suggest that optimized SLNs modified with 2-hydroxypropyl-β-cyclodextrin may have potential as an oral drug delivery system for PTX.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Antineoplastic Agents, Phytogenic; ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; beta-Cyclodextrins; Biological Transport; Breast Neoplasms; Calcium Channel Blockers; Cell Survival; Chemistry, Pharmaceutical; Drug Delivery Systems; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Excipients; Female; Humans; MCF-7 Cells; Nanoparticles; Neoplasm Proteins; Paclitaxel; Stearic Acids; Surface-Active Agents; Ultrasonics; Verapamil

Cancer cell migration is an early event in metastasis, the main cause of breast cancer-related deaths. Cholesterol-enriched membrane domains called lipid rafts influence the function of many molecules, including the raft-associated protein CD44. We describe a novel mechanism whereby rafts regulate interactions between CD44 and its binding partner ezrin in migrating breast cancer cells. Specifically, in nonmigrating cells, CD44 and ezrin localized to different membranous compartments: CD44 predominantly in rafts, and ezrin in nonraft compartments. After the induction of migration (either nonspecific or CD44-driven), CD44 affiliation with lipid rafts was decreased. This was accompanied by increased coprecipitation of CD44 and active (threonine-phosphorylated) ezrin-radixin-moesin (ERM) proteins in nonraft compartments and increased colocalization of CD44 with the nonraft protein, transferrin receptor. Pharmacological raft disruption using methyl-β-cyclodextrin also increased CD44-ezrin coprecipitation and colocalization, further suggesting that CD44 interacts with ezrin outside rafts during migration. Conversely, promoting CD44 retention inside lipid rafts by pharmacological inhibition of depalmitoylation virtually abolished CD44-ezrin interactions. However, transient single or double knockdown of flotillin-1 or caveolin-1 was not sufficient to increase cell migration over a short time course, suggesting complex crosstalk mechanisms. We propose a new model for CD44-dependent breast cancer cell migration, where CD44 must relocalize outside lipid rafts to drive cell migration. This could have implications for rafts as pharmacological targets to down-regulate cancer cell migration.

Topics: beta-Cyclodextrins; Breast Neoplasms; Caveolin 1; Cell Compartmentation; Cell Line, Tumor; Cell Movement; Cytoskeletal Proteins; Female; Gene Knockdown Techniques; Humans; Hyaluronan Receptors; Hyaluronic Acid; Membrane Microdomains; Membrane Proteins; Models, Biological; Protein Binding; Protein Transport; Subcellular Fractions

5-Fluorouracil (5-FU) in combination with its synergistic biomodulator folinic acid maintains a pivotal position in cancer chemotherapy. However, clinical limitations such as phlebitis and catheter blockages persist with the administration of these drugs in combination, and are associated with reduced efficacy and/or quality of life for patients. We have reported earlier on the novel, all-in-one, pH neutral, parenteral 5-FU and folinic acid formulations (termed Fluorodex) incorporating β-cyclodextrins. Fluorodex maintains potency while overcoming the accepted incompatibility of 5-FU and folinic acid. We carried out toxicological, pharmacokinetic and biodistribution, and efficacy evaluations of Fluorodex compared with 5-FU:folinic acid using several administration routes and schedules in two rodent models. These were compared with the dose-matched sequential administration of 5-FU:folinic acid. Fluorodex showed bioequivalence to 5-FU:folinic acid as assessed by the tissue distribution and pharmacokinetic studies of 5-FU, but was generally better tolerated as determined by weight loss, hematological, and other clinical parameters. Compared with 5-FU:folinic acid, Fluorodex was also associated with reduced phlebitis using a rabbit ear vein model. Furthermore, using human carcinoma tumor models in mice, Fluorodex resulted in equivalent or improved efficacy profiles compared with 5-FU:folinic acid. In conclusion, these novel, all-in-one formulations represent a superior injectable form of 5-FU that allows codelivery of folinic acid. This should translate into improved patient tolerability with potential for enhanced efficacy.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Colorectal Neoplasms; Dose-Response Relationship, Drug; Drug Combinations; Drug Screening Assays, Antitumor; Female; Fluorouracil; HCT116 Cells; HT29 Cells; Humans; Leucovorin; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Rabbits; Rats; Tissue Distribution; Xenograft Model Antitumor Assays

Beta cyclodextrin (β-CD) based nanosponges were synthesized and paclitaxel inclusion complex with nanosponges were prepared using techniques of inclusion complex formation. The paclitaxel nanosponge's complexes were evaluated for their release. The nanosponges complexes were also evaluated using DSC, FTIR, and NMR techniques for confirmation of inclusion complex formation between paclitaxel and nanosponges. Particle size and morphology of paclitaxel nanosponge's complex were estimated using SEM, TEM and dynamic light scattering techniques. The particle sizes were found out to be in range of 400 to 600 nm. Cytotoxic efficacy of paclitaxel nanosponge complex was determined against MCF-7 cells and paclitaxel nanosponge's complex was found to be cytotoxic and more effective against this cell line.

Topics: Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Calorimetry, Differential Scanning; Cell Line, Tumor; Cell Survival; Drug Carriers; Drug Compounding; Excipients; Female; Humans; Kinetics; Magnetic Resonance Spectroscopy; Nanostructures; Osmolar Concentration; Paclitaxel; Particle Size; Solubility; Spectroscopy, Fourier Transform Infrared

Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) exhibits potent antitumour activity via membrane receptors on cancer cells without deleterious side-effects for normal tissue. Unfortunately, like many other cancer types, breast cancer cells develop resistance to TRAIL; therefore, TRAIL-sensitising agents are currently being explored. In this study, we report that seleno-cyclodextrin (2-selenium-bridged β-cyclodextrin, 2-SeCD), a seleno-organic compound with glutathione peroxidase (GPx)-mimetic activity, sensitises TRAIL-resistant human breast cancer cells and xenograft tumours to undergo apoptosis. In vitro, 2-SeCD reduces the viability of cancer cells by inducing cell cycle arrest in G(2)/M phase. Furthermore, 2-SeCD efficiently sensitises MDA-MB-468 and T47D cells but not untransformed human mammary epithelial cells to TRAIL-mediated apoptosis, as evidenced by enhanced caspase activity and poly-ADP-ribose-polymerase (PARP) cleavage. From a mechanistic standpoint, we show that 2-SeCD induces the expression of TRAIL receptors DR5 but not DR4 on both mRNA and protein levels in a dose-dependent manner. Moreover, 2-SeCD treatment also suppresses TRAIL-induced nuclear factor-κB (NF-κB) pro-survival pathways by preventing cytosolic IκBα degradation and p65 nuclear translocation. Consequently, the combined administration suppresses anti-apoptotic proteins transcriptionally regulated by NF-κB. In vivo, 2-SeCD and TRAIL are well tolerated in mice, and their combination significantly inhibits the growth of MDA-MB-468 xenografts and promotes apoptosis. Up-regulation of DR5 and down-regulation of NF-κB by dual treatment were also observed in tumour tissues. Overall, 2-SeCD sensitises resistant breast cancer cells to TRAIL-based apoptosis in vitro and in vivo. These findings provide strong evidence for the therapeutic potential of this combination against breast cancers.

Topics: Animals; Antineoplastic Agents; Apoptosis; beta-Cyclodextrins; beta-Galactosidase; Blotting, Western; Breast Neoplasms; Caspases; Cell Cycle; Cell Line, Tumor; Cell Survival; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Luciferases; Mice; NF-kappa B; Organoselenium Compounds; Poly(ADP-ribose) Polymerases; Receptors, TNF-Related Apoptosis-Inducing Ligand; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; TNF-Related Apoptosis-Inducing Ligand; Transplantation, Heterologous

Epidermal growth factor receptor (EGFR) is overexpressed in many cancer types including ~30% of breast cancers. Several small molecule tyrosine kinase inhibitors (TKIs) targeting EGFR have shown clinical efficacy in lung and colon cancers, but no benefit has been noted in breast cancer. Thirteen EGFR expressing breast cancer cell lines were analyzed for response to EGFR TKIs. Seven were found to be EGFR TKI resistant; while shRNA knockdown of EGFR determined that four of these cell lines retained the requirement of EGFR protein expression for growth. Interestingly, EGFR localized to plasma membrane lipid rafts in all four of these EGFR TKI-resistant cell lines, as determined by biochemical raft isolation and immunofluorescence. When lipid rafts were depleted of cholesterol using lovastatin, all four cell lines were sensitized to EGFR TKIs. In fact, the effects of the cholesterol biosynthesis inhibitors and gefitinib were synergistic. While gefitinib effectively abrogated phosphorylation of Akt- and mitogen-activated protein kinase in an EGFR TKI-sensitive cell line, phosphorylation of Akt persisted in two EGFR TKI-resistant cell lines, however, this phosphorylation was abrogated by lovastatin treatment. Thus, we have shown that lipid raft localization of EGFR correlates with resistance to EGFR TKI-induced growth inhibition and pharmacological depletion of cholesterol from lipid rafts decreases this resistance in breast cancer cell lines. Furthermore, we have presented evidence to suggest that when EGFR localizes to lipid rafts, these rafts provide a platform to facilitate activation of Akt signaling in the absence of EGFR kinase activity.

Topics: Atorvastatin; Benzylamines; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cholesterol; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Drug Synergism; ErbB Receptors; Female; Gefitinib; Heptanoic Acids; Humans; Lovastatin; Membrane Microdomains; Phosphorylation; Protein Kinase Inhibitors; Protein Transport; Proto-Oncogene Proteins c-akt; Pyrroles; Quinazolines; Thiophenes

Exosomes are nano-vesicles secreted by a wide range of mammalian cell types. These vesicles are abundant in serum and other extracellular fluids and contain a large repertoire of proteins, mRNA and microRNA. Exosomes have been implicated in cell to cell communication, the transfer of infectious agents, and neurodegenerative diseases as well as tumor progression. However, the precise mechanisms by which they are internalized and/or secreted remain poorly understood. In order to follow their release and uptake in breast tumor cells in real time, cell-derived exosomes were tagged with green fluorescent protein (GFP)-CD63 while human serum exosomes were rhodamine isothiocynate-labeled. We show that detachment of adherent cells from various substrata induces a rapid and substantial secretion of exosomes, which then concentrate on the cell surfaces and mediate adhesion to various extracellular matrix proteins. We also demonstrate that disruption of lipid rafts with methyl-beta-cyclodextrin (MβCD) inhibits the internalization of exosomes and that annexins are essential for the exosomal uptake mechanisms. Taken together, these data suggest that cellular detachment is accompanied by significant release of exosomes while cellular adhesion and spreading are enhanced by rapid uptake and disposition of exosomes on the cell surface.

Topics: Annexins; beta-Cyclodextrins; Biological Transport; Blotting, Western; Breast Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Movement; Exosomes; Female; Flow Cytometry; Fluorescence Recovery After Photobleaching; Humans

Adjuvant treatment with tamoxifen substantially improves survival of women with estrogen-receptor positive (ER+) tumors. Tamoxifen resistance (TAMR) limits clinical benefit. RRR-α-tocopherol ether-linked acetic acid analogue (α-TEA) is a small bioactive lipid with potent anticancer activity. We evaluated the ability of α-TEA in the presence of tamoxifen to circumvent TAMR in human breast cancer cell lines.. Two genotypically matched sets of TAM-sensitive (TAMS) and TAM-resistant (TAMR) human breast cancer cell lines were assessed for signal-transduction events with Western blotting, apoptosis induction with Annexin V-FITC/PI assays, and characterization of cholesterol-rich microdomains with fluorescence staining. Critical involvement of selected mediators was determined by using RNA interference and chemical inhibitors.. Growth-factor receptors (total and phosphorylated forms of HER-1 and HER-2), their downstream prosurvival mediators pAkt, pmTOR, and pERK1/2, phosphorylated form of estrogen receptor-α (pER-α at Ser-167 and Ser-118, and cholesterol-rich lipid microdomains were highly amplified in TAMR cell lines and enhanced by treatment with TAM. α-TEA disrupted cholesterol-rich microdomains, acted cooperatively with TAM to reduce prosurvival mediators, and induced DR5-mediated mitochondria-dependent apoptosis via an endoplasmic reticulum stress-triggered pro-death pJNK/CHOP/DR5 amplification loop. Furthermore, methyl-β-cyclodextrin (MβCD), a chemical disruptor of cholesterol rich microdomains, acted cooperatively with TAM to reduce prosurvival mediators and to induce apoptosis.. Data for the first time document that targeting cholesterol-rich lipid microdomains is a potential strategy to circumvent TAMR, and the combination of α-TEA + TAM can circumvent TAMR by suppression of prosurvival signaling via disruption of cholesterol-rich lipid microdomains and activation of apoptotic pathways via induction of endoplasmic reticulum stress.

Topics: alpha-Tocopherol; Antineoplastic Agents, Hormonal; Antioxidants; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Cholesterol; Drug Resistance, Neoplasm; Female; Humans; Membrane Microdomains; Signal Transduction; Tamoxifen

Topics: Animals; Antimetabolites, Antineoplastic; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chemistry, Pharmaceutical; Chromatography, Gel; Dendrimers; Dose-Response Relationship, Drug; Drug Carriers; Drug Compounding; Female; Humans; Inhibitory Concentration 50; Kinetics; Lysosomes; Magnetic Resonance Spectroscopy; Methotrexate; Micelles; Microscopy, Fluorescence; Particle Size; Polyethylene Glycols; Solubility; Technology, Pharmaceutical

Oxidative stress has emerged as an important pathogenic factor in the development of breast cancer. Cholesterol-rich membrane rafts or lipid rafts (LRs) are reported to play an important role in oxidative stress-induced signal transduction. NADPH oxidase-dependent reactive oxygen species (ROS) production is implicated in oxidative stress in human mammary epithelial cells. In the present study, we determined the expression and regulation of membrane-bound subunits by LRs in human breast cancer cells. We report that basal levels of gp91phox and p22phox are expressed in breast cancer cells. We demonstrate for the first time that disruption of LRs resulted in the downregulation of NADPH oxidase subunits in breast cancer cells. Cholesterol depletion by 10 mM methyl-β-cyclodextrin (MβCD) translocated both gp91phox and p22phox out of LRs. Moreover, lipid raft disruption decreased NADPH oxidase activity (21.1 ± 0.5% in MCF-7 and 28.9 ± 1.0 in BT-549 cells), which was reversed by cholesterol repletion (95%). Therefore, the results suggest that the integrity of LRs plays an important role in the regulation of NADPH oxidase activity in breast cancer cells.

Topics: beta-Cyclodextrins; Breast Neoplasms; Carcinoma; Cell Line, Tumor; Dose-Response Relationship, Drug; Down-Regulation; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Membrane Glycoproteins; Membrane Microdomains; NADPH Oxidase 2; NADPH Oxidases

uPAR and MMP-9, which play critical roles in tumor cell invasion, migration and angiogenesis, have been shown to be associated with lipid rafts.. To investigate whether cholesterol could regulate uPAR and MMP-9 in breast carcinoma, we used MβCD (methyl beta cyclodextrin, which extracts cholesterol from lipid rafts) to disrupt lipid rafts and studied its effect on breast cancer cell migration, invasion, angiogenesis and signaling.. Morphological evidence showed the association of uPAR with lipid rafts in breast carcinoma cells. MβCD treatment significantly reduced the colocalization of uPAR and MMP-9 with lipid raft markers and also significantly reduced uPAR and MMP-9 at both the protein and mRNA levels. Spheroid migration and invasion assays showed inhibition of breast carcinoma cell migration and invasion after MβCD treatment. In vitro angiogenesis studies showed a significant decrease in the angiogenic potential of cells pretreated with MβCD. MβCD treatment significantly reduced the levels of MMP-9 and uPAR in raft fractions of MDA-MB-231 and ZR 751 cells. Phosphorylated forms of Src, FAK, Cav, Akt and ERK were significantly inhibited upon MβCD treatment. Increased levels of soluble uPAR were observed upon MβCD treatment. Cholesterol supplementation restored uPAR expression to basal levels in breast carcinoma cell lines. Increased colocalization of uPAR with the lysosomal marker LAMP1 was observed in MβCD-treated cells when compared with untreated cells.. Taken together, our results suggest that cholesterol levels in lipid rafts are critical for the migration, invasion, and angiogenesis of breast carcinoma cells and could be a critical regulatory factor in these cancer cell processes mediated by uPAR and MMP-9.

Topics: beta-Cyclodextrins; Breast Neoplasms; Caveolins; Cell Line, Tumor; Cell Movement; Cholesterol; Culture Media, Conditioned; Dose-Response Relationship, Drug; Endothelial Cells; Extracellular Signal-Regulated MAP Kinases; Female; Focal Adhesion Kinase 1; Gene Expression Regulation; Humans; Matrix Metalloproteinase 9; Membrane Microdomains; Neoplasm Invasiveness; Neovascularization, Pathologic; Nystatin; Phosphorylation; Proto-Oncogene Proteins c-akt; Receptors, Urokinase Plasminogen Activator; RNA, Messenger; src-Family Kinases; Time Factors

We have previously shown that modification of polyphenols with a ferrocenyl group can dramatically enhance their cytotoxicity. We now present two new [3]ferrocenophane compounds, one of which has an antiproliferative effect seven times stronger than the corresponding noncyclic species, with IC50 values of 90 and 94 nM on hormone-independent MDA-MB-231 breast and PC-3 prostate cancer cell lines, respectively. Solubility studies in water using methylated beta-cyclodextrin and electron transfer studies are also presented.

Topics: Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Female; Flavonoids; Humans; Male; Phenols; Polyphenols; Prostatic Neoplasms; Structure-Activity Relationship

Recently we have described an HPMA copolymer conjugate carrying both the aromatase inhibitor aminoglutethimide (AGM) and doxorubicin (Dox) as combination therapy. This showed markedly enhanced in vitro cytotoxicity compared to the HPMA copolymer-Dox (FCE28068), a conjugate that demonstrated activity in chemotherapy refractory breast cancer patients during early clinical trials. To better understand the superior activity of HPMA copolymer-Dox-AGM, here experiments were undertaken using MCF-7 and MCF-7ca (aromatase-transfected) breast cancer cell lines to: further probe the synergistic cytotoxic effects of AGM and Dox in free and conjugated form; to compare the endocytic properties of HPMA copolymer-Dox-AGM and HPMA copolymer-Dox (binding, rate and mechanism of cellular uptake); the rate of drug liberation by lysosomal thiol-dependant proteases (i.e. conjugate activation), and also, using immunocytochemistry, to compare their molecular mechanism of action. It was clearly shown that attachment of both drugs to the same polymer backbone was a requirement for enhanced cytotoxicity. FACS studies indicated both conjugates have a similar pattern of cell binding and endocytic uptake (at least partially via a cholesterol-dependent pathway), however, the pattern of enzyme-mediated drug liberation was distinctly different. Dox release from PK1 was linear with time, whereas the release of both Dox and AGM from HPMA copolymer-Dox-AGM was not, and the initial rate of AGM release was much faster than that seen for the anthracycline. Immunocytochemistry showed that both conjugates decreased the expression of ki67. However, this effect was more marked for HPMA copolymer-Dox-AGM and, moreover, only this conjugate decreased the expression of the anti-apoptotic protein bcl-2. In conclusion, the superior in vitro activity of HPMA copolymer-Dox-AGM cannot be attributed to differences in endocytic uptake, and it seems likely that the synergistic effect of Dox and AGM is due to the kinetics of intracellular drug liberation which leads to enhanced activity.

Topics: Aminoglutethimide; Antineoplastic Agents; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Survival; Chlorpromazine; Cytochalasin B; Doxorubicin; Endocytosis; Female; Flow Cytometry; Humans; Immunohistochemistry; Ki-67 Antigen; Kinetics; Liver; Lysosomes; Methacrylates; Microscopy, Confocal; Proto-Oncogene Proteins c-bcl-2; Tetrazolium Salts; Thiazoles

In the following experiments, we sought to understand the triggering mechanism which propels galectin-3 to be secreted into the extracellular compartment from its intracellular stores in breast carcinoma cells. We also wanted to analyze in greater details the role of galectin-3 in cellular adhesion and spreading. To do this, we made use of two pairs of breast carcinoma cell lines where one of the pair has high expression of galectin-3 and the other low expression of the lectin. We determined that galectin-3 secreted into the conditioned medium of sub-confluent and spread cells in culture was quite low, almost negligible. However, once the cells were detached and rounded up, a mechano-sensing mechanism triggered the rapid secretion of galectin-3 into the conditioned medium. The secretion was constitutive as long as the cells remained detached. Galectin-3 was shown to be actively taken up from the conditioned medium by spreading cells. The cells which express and secrete high levels of galectin-3 adhered and spread much faster on plastic than those with reduced expression. The uptake of galectin-3 according to our data was important in cell spreading because if this process was compromised significantly, cells failed to spread. The data suggested that galectin-3 uptake modulates the adhesion plaques in that cells which express high levels of galectin-3 have thin-dot like plaques that may be suited for rapid adhesion and spreading while cells in which galectin-3 expression is reduced or knocked-down, have thick and elongated plaques which may be suited for a firmer adhesion to the substratum. Recombinant galectin-3 added exogenously reduced the thickness of the adhesion plaques of tumor cells with reduced galectin-3 expression. Taken together, the present data suggest that galectin-3 once externalized, is a powerful modulator of cellular adhesion and spreading in breast carcinoma cells.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; beta-Cyclodextrins; Breast Neoplasms; Carcinoma; Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Cells, Cultured; Extracellular Space; Focal Adhesions; Galectin 3; Humans; Mechanotransduction, Cellular; Protein Transport

Lipid rafts/caveolae are membrane platforms for signaling molecules that regulate various cellular functions, including cell survival. To better understand the role of rafts in tumor progression and therapeutics, we investigated the effect of raft disruption on cell viability and compared raft levels in human cancer cell lines versus their normal counterparts. Here, we report that cholesterol depletion using methyl-beta cyclodextrin caused anoikis-like apoptosis, which in A431 cells involved decreased raft levels, Bcl-xL down-regulation, caspase-3 activation, and Akt inactivation regardless of epidermal growth factor receptor activation. Cholesterol repletion replenished rafts on the cell surface and restored Akt activation and cell viability. Moreover, the breast cancer and the prostate cancer cell lines contained more lipid rafts and were more sensitive to cholesterol depletion-induced cell death than their normal counterparts. These results indicate that cancer cells contain increased levels of rafts and suggest a potential use of raft-modulating agents as anti-cancer drugs.

Topics: Apoptosis; bcl-X Protein; beta-Cyclodextrins; Breast Neoplasms; Carcinoma, Squamous Cell; Caspase 3; Caspases; Cell Line, Tumor; Cell Survival; Cholesterol; Down-Regulation; Enzyme Activation; ErbB Receptors; Female; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Male; Membrane Microdomains; Neoplasms; Prostatic Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; Simvastatin

The response rates of extensively used chemotherapeutic drugs, carboplatin (Carb) or 5-fluorouracil (5-FU) are relatively disappointing because of considerable side effects associated with their high-dose regimen. In the present study, we determined whether treatment with a cholesterol depleting agent, methyl-beta-cyclodextrin (MCD), enhances the weak efficacy of low doses of Carb or 5-FU in human breast cancer cells. Data demonstrate that pretreatment with MCD significantly potentiates the cytotoxic activity of Carb and 5-FU in both MCF-7 and MDA-MB-231. Furthermore, we explored the molecular basis of enhanced cytotoxicity, and our data revealed that low-dose treatment with these drugs in MCD pretreated cells exhibited significantly decreased Akt phosphorylation, NF-kappaB activity and down-regulation in expression of anti-apoptotic protein Bcl-2. In addition, MCD pretreated cells demonstrated an increased intracellular drug accumulation as compared to cells treated with drugs alone. Taken together, our data provide the basis for potential therapeutic application of MCD in combination with other conventional cytotoxic drugs to facilitate reduction of drug dosage that offers a better chemotherapeutic approach with low toxicity.

Topics: Adjuvants, Pharmaceutic; Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Carboplatin; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Down-Regulation; Drug Combinations; Drug Screening Assays, Antitumor; Female; Fluorouracil; Humans; NF-kappa B; Oncogene Protein v-akt; Phosphorylation; Proto-Oncogene Proteins c-bcl-2

In this report we show, by confocal analysis of indirect immunofluorescence, that the type-1 cannabinoid receptor (CB1R), which belongs to the family of G-protein-coupled receptors, is expressed on the plasma membrane in human breast cancer MDA-MB-231 cells. However, a substantial proportion of the receptor is present in lysosomes. We found that CB1R is associated with cholesterol- and sphyngolipid-enriched membrane domains (rafts). Cholesterol depletion by methyl-beta-cyclodextrin (MCD) treatment strongly reduces the flotation of the protein on the raft-fractions (DRM) of sucrose density gradients suggesting that CB1 raft-association is cholesterol dependent. Interestingly binding of the agonist, anandamide (AEA) also impairs DRM-association of the receptor suggesting that the membrane distribution of the receptor is dependent on rafts and is possibly regulated by the agonist binding. Indeed MCD completely blocked the clustering of CB1R at the plasma membrane. On the contrary the lysosomal localization of CB1R was impaired by this treatment only after AEA binding.

Topics: Arachidonic Acids; beta-Cyclodextrins; Breast Neoplasms; Cell Line, Tumor; Cell Membrane; Cholesterol; Endocannabinoids; Humans; Lysosomes; Membrane Microdomains; Polyunsaturated Alkamides; Receptor, Cannabinoid, CB1

Methyl-beta-cyclodextrin (MEBCD) was investigated for its effect on the antitumoral activity of various antineoplastic agents (doxorubicin (DOX), docetaxel (DXL), 5-fluorouracil (5-FU) and cisplatin (CDDP)) in three different human parental sensitive cancer cell lines (K562 S, MCF7 S and A2780 S) and their multidrug resistant variant sublines (K562 R, MCF7 R and A2780 R). At non-cytotoxic concentrations, MEBCD was able to increase significantly DOX and DXL cytotoxic activity in all the cell lines tested. The sensitisation ratios (IC50 drug control/IC50 drug-MEBCD treated) ranged from 3l1 to 14.3. Moreover, intracellular DOX accumulation, determined by high-performance liquid chromatography, was also increased when cells were treated with MEBCD combined with DOX (approximately 2-3 fold). The effects of MEBCD in resistant sublines were greater than in their parental sensitive cell lines. Other experiments demonstrated that the action of the MEBCD was independent of DOX. These data provided a basis for the potential therapeutic application of MEBCD in cancer therapy.

Topics: Antibiotics, Antineoplastic; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Breast Neoplasms; Cisplatin; Cyclodextrins; Docetaxel; Doxorubicin; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Synergism; Female; Fluorouracil; Humans; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Ovarian Neoplasms; Paclitaxel; Taxoids; Tumor Cells, Cultured

The anti-tumour activity of methyl-beta-cyclodextrin (MEBCD), a cyclic oligosaccharide known for its interaction with the plasma membrane, was investigated in vitro and in vivo and compared with that of doxorubicin (DOX) in the human tumour models MCF7 breast carcinoma and A2780 ovarian carcinoma. In vitro proliferation was assessed using the MTT assay. In vivo studies were carried out using xenografted Swiss nude mice injected weekly i.p. with MEBCD at 300 or 800 mg kg(-1) or DOX at 2 mg kg(-1), during 2 months. Under these conditions, MEBCD was active against MCF7 and A2780 cell lines and tumour xenografts. For each tumour model, the tumoral volume of the xenografted mice treated with MEBCD was at least twofold reduced compared with the control group. In the MCF7 model, MEBCD (800 mg kg(-1)) was more active than DOX (2 mg kg(-1)). After 56 days of treatment with MEBCD, no toxicologically meaningful differences were observed in macroscopic and microscopic parameters compared with controls. The accumulation of MEBCD in normal and tumour tissues was also assessed using a chromatographic method. Results indicated that after a single injection of MEBCD, tumour, liver and kidneys accumulated the highest concentrations of MEBCD. These results provided a basis for the potential therapeutic application of MEBCD in cancer therapy.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; beta-Cyclodextrins; Breast Neoplasms; Cell Division; Cyclodextrins; Female; Humans; Lethal Dose 50; Mice; Mice, Nude; Neoplasm Transplantation; Ovarian Neoplasms; Tissue Distribution; Transplantation, Heterologous; Tumor Cells, Cultured