curcumin and thiazolyl-blue

The present work involves encapsulation of herbal drug nanocurcumin into the virosomes and compared with a liposome in terms of their in vitro anti-proliferative, anti-inflammatory, and anti-migratory efficacy.. The anti-proliferative, anti-inflammatory, and anti-migratory efficacy of virosome and liposome were compared in HepG2 and CaCo2 cells by using MTT, Nitric oxide scavenging, and Wound healing assay, respectively.. Size of the optimised NC-Virosome and NC-Liposome was 70.06 ± 1.63 and 265.80 ± 1.64 nm, respectively. The prepared NC-Virosome can be stored at -4 °C up to six months. The drug encapsulation efficiency of NC-Virosome and NC-Liposome was found to be 84.66 ± 1.67 and 62.15 ± 1.75% (w/w). The evaluated minimum inhibitory concentration (IC50 value) for NC-Virosome was 102.7 μg/ml and 108.1 μg/ml, while NC-Liposome showed 129.2 μg/ml and 160.1 μg/ml for HepG2 and CaCo2 cells, respectively. Morphological examination depicts detachment of the cells from substratum after exposure to NC-Virosome for 48 h.. The prepared NC-Virosome provides remarkable in vitro efficacy in both the cell lines with site-specific drug-targeting potential as compared to the liposome, results proved its potential as a drug delivery vehicle for future therapy with reduced toxicity.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents, Phytogenic; Caco-2 Cells; Cell Movement; Curcumin; Drug Delivery Systems; Excipients; Hep G2 Cells; Humans; Liposomes; Microbial Sensitivity Tests; Neoplasms; Nitric Oxide; Tetrazolium Salts; Thiazoles; Virosomes; Wound Healing

To develop docetaxel (DT) and curcumin (CUR) co-loaded nanostructured lipid carriers (DTCR-NLCs) for ratiometric co-targeting to non-small cell lung carcinoma (NSCLC) cells.. The DTCR-NLCs were developed by employing a high-pressure homogenisation technique and optimised by employing a rotatable central composite design response surface methodology (RCCD-RSM) via the design of experiments (DoE) approach.. The optimised DTCR-NLCs had a particle size (D90) of 150.2 ± 5.2 nm, Pdi of 0.263 ± 0.15, zeta potential of +26.3 ± 5.2 mv. The % drug loading (% DL) of DT and CUR was observed to be 1.38 ± 0.98 and 2.99 ± 1.24, respectively. Dissolution studies depicted a pH-independent drug release (≈98% drug release at 144 h). The DTCR-NLCs were stable and haemocompatible. MTT cell viability assay of DTCR-NLCs demonstrated considerably increased cytotoxicity towards NCI-H460 cells.. The developed DTCR-NLCs heralds the future of an efficacious and safer Taxane therapy for NSCLC.

Topics: Antineoplastic Agents; Calorimetry, Differential Scanning; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Survival; Cobalt; Curcumin; Docetaxel; Drug Delivery Systems; Drug Screening Assays, Antitumor; Hemolysis; Humans; Hydrogen-Ion Concentration; In Vitro Techniques; Lipids; Lung Neoplasms; Microspheres; Nanostructures; Particle Size; Temperature; Tetrazolium Salts; Thiazoles; X-Ray Diffraction

Curcumin, a major component of curry powder, which is a natural polyphenol product extracted from rhizoma curcumae longae, interacts with a specific binding site on microtubules. Vinblastine is an antitumor drug that induces microtubule depolymerization.. We investigated whether curcumin influences the antitumor effect of vinblastine in HeLa human cervical cancer cells.. Changes in microtubule filaments were visualized by immuno-staining. Cell death was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) or water-soluble tetrazolium(WST) assay. Apoptotic cell formation was assessed by flow cytometry after staining cells with propidium iodide(PI) and/or Annexin V or with 6-diamidino-2-phenylindole(DAPI). Reactive oxygen species(ROS) were also measured by flow cytometry using dichloro-dihydro-fluorescein diacetate(DCF-DA). JC-1 was used to determine mitochondrial membrane potential (MMP).. When cells were pretreated with curcumin, microtubule filaments were disordered. Vinblastine-induced microtubule depolymerization and cell death were reduced in HeLa human cervical cancer cells pretreated with curcumin compared to the control. The decrease in cell death was much greater in cells pretreated with curcumin compared to cotreatment or post-treatment. DNA condensation by vinblastine was also decreased in curcumin-pretreated cells. Curcumin reduced ROS production by vinblastine. However, no changes in vinblastine-mediated microtubule depolymerization were detected upon N-acetylcysteine(NAC) treatment. In contrast, vinblastine-induced MMP collapse was inhibited by pretreatment with curcumin or NAC. These findings suggest that vinblastine-induced tumor cell death might be inhibited by curcumin via ROS-independent microtubule dynamics and ROS-dependent MMP collapse. It also suggests that microtubule dynamics could be necessary for the optimal antitumor activity of vinblastine. Our results suggest that patients treated with vinblastine should not consume curcumin.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Caspase 3; Cell Survival; Curcumin; Drug Synergism; Female; HeLa Cells; Humans; Male; Membrane Potential, Mitochondrial; Mice, Inbred C57BL; Microtubules; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles; Vinblastine

Pulp repair is less likely to occur when dentin or pulpal tissue remains infected after caries excavation. Yet there are currently few options to kill residual bacteria without damaging resident cells. The current study has evaluated the effect of 3 blue light-activated chemicals on the viability of lactobacilli, odontoblast-like cells (MDPC-23), undifferentiated pulp cells (OD21), and human embryonic stem cells (hESC H1).. Bacteria were incubated for 15 minutes with curcumin, eosin Y, or rose bengal and then irradiated with blue light (240 seconds). Bacteria were labeled with LIVE/DEAD BacLight Bacterial Viability kit, and viability was assessed by fluorescence-activated cell sorting. Cytotoxicity assays were performed on MDPC-23 cells, OD21, and hESC H1 cells grown in 24-well plates and exposed to the same photosensitizer-light combination. After 24 hours, cellular response was measured by using the methyl-thiazol-diphenyl-tetrazolium assay. Results were statistically analyzed by using one-way analysis of variance and Tukey multiple comparison intervals (α = 0.05).. Bacterial viability was significantly reduced after exposure to different combinations of light and photosensitizers; mitochondrial activity of cultured cells remained unaffected when exposed to the same conditions, suggesting a good therapeutic index in vitro.. Blue light-mediated disinfection is promising for the development of new treatment strategies designed to promote pulp repair after carious exposure.

Topics: Animals; Anti-Infective Agents; Cell Line; Cell Survival; Coloring Agents; Curcumin; Dental Pulp; Embryonic Stem Cells; Eosine Yellowish-(YS); Humans; Lactobacillus; Light; Mice; Microbial Viability; Mitochondria; Odontoblasts; Photosensitizing Agents; Rose Bengal; Tetrazolium Salts; Thiazoles

Curcumin is a multi-targeted anti-cancer agent. However, there are few studies on its anti-leukemia activity in human acute monocytic leukemia. Here, we study the effect and mechanisms of curcumin on acute monocytic leukemia.. The acute monocytic leukemia cell line THP-1 was used as in vitro cell model to explore the anti-leukemia effects and mechanisms of curcumin. Cell proliferation was measured by MTT assay, cell apoptosis bodies were observed using a light microscope, cell apoptosis rate was evaluated by flow cytometry, and the expression alterations of growth-sinaling proteins were detected by Western blotting.. Curcumin inhibited cell proliferation and induced cell apoptosis in time- and dose- dependent manner in THP-1 cells. Curcumin significantly inhibited the activations of AKT/mTOR and RAF/MEK/ERK signaling pathways simultaneously.. This study demonstrates that curcumin inhibits proliferation and induces apoptosis in THP-1 cells via inhibiting the activations of AKT/mTOR and RAF/MEK/ERK signaling pathways simultaneously. Our data suggest that curcumin is a promising anti-tumor agent in acute monocytic leukemia.

Topics: Annexin A5; Antineoplastic Agents; Apoptosis; Blotting, Western; Cell Line, Tumor; Cell Proliferation; Coloring Agents; Curcumin; Enzyme Activation; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Humans; Leukemia, Myeloid, Acute; MAP Kinase Kinase Kinases; MAP Kinase Signaling System; Oncogene Protein v-akt; raf Kinases; Tetrazolium Salts; Thiazoles; TOR Serine-Threonine Kinases

Curcuma zedoaria also known as Temu putih is traditionally used in food preparations and treatment of various ailments including cancer. The cytotoxic activity of hexane, dichloromethane, ethyl acetate, methanol, and the methanol-soxhlet extracts of Curcuma zedoaria rhizomes was tested on two human cancer cell lines (Ca Ski and MCF-7) and a noncancer cell line (HUVEC) using MTT assay. Investigation on the chemical components in the hexane and dichloromethane fractions gave 19 compounds, namely, labda-8(17),12 diene-15,16 dial (1), dehydrocurdione (2), curcumenone (3), comosone II (4), curcumenol (5), procurcumenol (6), germacrone (7), zerumbone epoxide (8), zederone (9), 9-isopropylidene-2,6-dimethyl-11-oxatricyclo[6.2.1.0(1,5)]undec-6-en-8-ol (10), furanodiene (11), germacrone-4,5-epoxide (12), calcaratarin A (13), isoprocurcumenol (14), germacrone-1,10-epoxide (15), zerumin A (16), curcumanolide A (17), curcuzedoalide (18), and gweicurculactone (19). Compounds (1-19) were evaluated for their antiproliferative effect using MTT assay against four cancer cell lines (Ca Ski, MCF-7, PC-3, and HT-29). Curcumenone (3) and curcumenol (5) displayed strong antiproliferative activity (IC50 = 8.3 ± 1.0 and 9.3 ± 0.3 μg/mL, resp.) and were found to induce apoptotic cell death on MCF-7 cells using phase contrast and Hoechst 33342/PI double-staining assay. Thus, the present study provides basis for the ethnomedical application of Curcuma zedoaria in the treatment of breast cancer.

Topics: Analysis of Variance; Breast Neoplasms; Cell Proliferation; Chromatography, Thin Layer; Curcuma; Female; Human Umbilical Vein Endothelial Cells; Humans; Indonesia; Malaysia; MCF-7 Cells; Microscopy, Fluorescence; Molecular Structure; Phytotherapy; Plant Extracts; Rhizome; Tetrazolium Salts; Thiazoles

Signal transducer and activator of transcription 3 (STAT3) is an oncogenic transcription factor constitutively active and aberrantly expressed in cervical cancer. However, the functional role of STAT3 in regulation of HPV's viral oncogene expression and downstream events associated with cervical carcinogenesis is not known. Our present study performed on HPV16-positive cervical cancer cell lines (SiHa and CaSki) and primary tumor tissues revealed a strong positive correlation of constitutively active STAT3 with expression of HPV16 E6 and E7 oncoproteins and a negative association with levels of p53 and pRB. Pharmacologic targeting of STAT3 expression in cervical cancer cell lines either by STAT3-specific siRNA or blocking its tyrosine phosphorylation by AG490 or curcumin led to dose-dependent accumulation of p53 and pRb in cervical cancer cells. Interestingly, the suppression of STAT3 expression or activation was associated with the gradual loss of HPV16 E6 and E7 expression and was accompanied by loss of cell viability. The viability loss was specifically high in HPV16-positive cells as compared to HPV negative C33a cells. These findings substantiate the regulatory role of STAT3 in HPV16-mediated cervical carcinogenesis. Leads obtained from the present study provide a strong rationale for developing novel STAT3-based approaches for therapeutic interventions against HPV infection to control cervical cancer.

Topics: Carcinogenesis; Caspase 3; Cell Line, Tumor; Curcumin; Electrophoretic Mobility Shift Assay; Female; Flow Cytometry; Gene Expression Regulation, Viral; Human papillomavirus 16; Humans; Immunoblotting; Immunohistochemistry; Oncogene Proteins, Viral; RNA, Small Interfering; STAT3 Transcription Factor; Tetrazolium Salts; Thiazoles; Tyrphostins; Uterine Cervical Neoplasms

Wound healing is a complex process involving many interdependent and overlapping sequences of physiological actions. The application of exogenous lactate released from poly (lactic-co-glycolic acid) (PLGA) polymer accelerated angiogenesis and wound healing processes. Curcumin is a well-known topical wound healing agent for both normal and diabetic-impaired wounds. Hence, we hypothesized that the PLGA nanoparticles encapsulating curcumin could much potentially accelerate the wound healing. In a full thickness excisional wound healing mouse model, PLGA-curcumin nanoparticles showed a twofold higher wound healing activity compared to that of PLGA or curcumin. Histology and RT-PCR studies confirmed that PLGA-curcumin nanoparticles exhibited higher re-epithelialization, granulation tissue formation and anti-inflammatory potential. PLGA nanoparticles offered various benefits for the encapsulated curcumin like protection from light degradation, enhanced water solubility and showed a sustained release of curcumin over a period of 8 days. In conclusion, we demonstrated the additive effect of lactic acid from PLGA and encapsulated curcumin for the active healing of wounds.

Topics: Animals; Anti-Inflammatory Agents; Cell Line, Tumor; Cell Survival; Collagen; Curcumin; Female; Glutathione Peroxidase; Keratinocytes; L-Lactate Dehydrogenase; Lactic Acid; Mice; Nanoparticles; NF-kappa B; Peroxidase; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Skin; Tetrazolium Salts; Thiazoles; Wound Healing

The unfolded protein response, autophagy and endoplasmic reticulum (ER) stress-induced apoptosis regulate tumor cell fate and have become novel signaling targets for the development of cancer therapeutic drugs. Curcumin has been used to treat several different cancers, including ovarian cancer, in clinical trials and research; however, the role of ER stress and autophagy in the therapeutic effects of curcumin and new curcumin analogues remains unclear.. Cell viability was determined using the MTT assay. Apoptosis was detected using flow cytometry with PI/Annexin V-FITC staining. The expression levels of ER stress- and autophagy-related proteins were analyzed by western blotting. The activation of autophagy was detected using immunofluorescence staining.. We demonstrated that B19 induced HO8910 cell apoptosis in a dose-responsive manner. We also determined and that this effect was associated with corresponding increases in a series of key components in the UPR and ER stress-mediated apoptosis pathways, followed by caspase 3 cleavage and activation. We also observed that B19 treatment induced autophagy in HO8910 cells. The inhibition of autophagy using 3-methyladenine (3-MA) increased levels of intracellular misfolded proteins, which enhanced ovarian cancer apoptosis.. Our data indicate that ER stress and autophagy may play a role in the apoptosis that is induced by the curcumin analogue B19 in an epithelial ovarian cancer cell line and that autophagy inhibition can increase curcumin analogue-induced apoptosis by inducing severe ER stress.

Topics: Analysis of Variance; Apoptosis; Autophagy; Blotting, Western; Cell Line, Tumor; Curcumin; Dose-Response Relationship, Drug; Endoplasmic Reticulum Stress; Female; Flow Cytometry; Fluorescent Antibody Technique; Humans; Microscopy, Electron; Signal Transduction; Tetrazolium Salts; Thiazoles

An in vitro study was made to evaluate the effect of curcumin and irradiation upon oral squamous cell carcinoma.. Curcumin was administered at doses of 3, 3.75, 4.50 and 5.25 μM in PE/CA-PJ15 oral squamous cell carcinoma cultures irradiated with different doses (1, 2.5 and 5 Gy), followed by evaluation of the effects upon cell viability after 24, 48 and 72 h, based on the MTT colorimetric test.. The application of curcumin to the PECA/PJ15 tumor cells during 24, 48 and 72 h of incubation without irradiation exerted an inhibitor effect upon cell viability. The curcumin concentration at which the inhibition of cell viability proved maximum was 5.25 μM, with statistically significant differences for 24 h (p = 0.002), 48 h (p < 0.001) and 72 h of incubation (p < 0.001). In contrast, the combination of curcumin and irradiation exerted a synergic effect-the greatest effects in relation to cell viability being recorded with a curcumin concentration of 3.75 μM and 5 Gy of irradiation, in the studied cell line.. Curcumin increases cytotoxic activity in the PE/CA PJ15 cell line, while the combination of curcumin and irradiation exerts a synergic effect.

Topics: Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Survival; Colorimetry; Coloring Agents; Curcumin; Humans; Mouth Neoplasms; Neoadjuvant Therapy; Radiation-Sensitizing Agents; Radiotherapy Dosage; Tetrazolium Salts; Thiazoles; Time Factors

Aluminum is neurotoxic both in animals and human beings primarily because of its interference with biological enzymes in key mechanisms of metabolic pathways. Mitochondrial dysfunction is one such mechanism that has been implicated in the pathogenesis of neurodegenerative diseases like Alzheimer's disease. Aluminum toxicity is very closely related to Alzheimer's disease. We evaluated the potentials of curcumin, a known cytoprotectant, against neurotoxic consequences of aluminum that acts through a wide range of mechanisms. Curcumin has been reported to be an antioxidant, and it is this property that is widely held to be responsible for its protective effects in tissue. Aluminum was administered by oral gavage at a dose level of 100 mg/kg body wt/day for a period of 8 weeks. Curcumin was administered in conjunction with aluminum at a dose of 50 mg/kg of body wt i.p. for a period of 8 weeks on alternate days. The effects of different treatments were studied on oxidative phosphorylation and reduced glutathione of different regions of rat brain. The study indicates reduced activity of NADH dehydrogenase (complex I), succinic dehydrogenase (complex II), and cytochrome oxidize (Complex IV) in all the three regions of rat brain, i.e., cerebral cortex, mid brain, and cerebellum. Curcumin supplementation to aluminum-treated rats was able to normalize significantly the activities of all the three mitochondrial complexes as well as reduced glutathione content in all the three regions of brain which were altered following aluminum treatment. We conclude that curcumin, by attenuating oxidative stress, as evident by hypoxia in histological observations and mitochondrial dysfunction holds a promise as an agent that can potentially reduce aluminum-induced adverse effects in brain.

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Aluminum; Animals; Anti-Inflammatory Agents, Non-Steroidal; Brain; Curcumin; Electron Transport Complex IV; Gene Expression Regulation; Glutathione; HSP72 Heat-Shock Proteins; Mitochondria; NADH Dehydrogenase; Oxidative Stress; Rats; Rats, Sprague-Dawley; RNA, Messenger; Succinate Dehydrogenase; Tetrazolium Salts; Thiazoles

Multidrug resistance (MDR) continues to be a major obstacle for successful anticancer therapy. In this work, fractions from 17 clinically used antitumour traditional Chinese medicinal herbs were tested for their potential to restore the sensitivity of MCF-7/ADR and A549/Taxol cells to a known antineoplastic agent. The effects of these fractions were evaluated by MTT method and an assay of the cellular accumulation of doxorubicin. Fractions from the PB group (herbs with the ability to promote blood circulation and remove blood stasis) showed more significant effects than fractions from the CH group (herbs with the ability to clear away heat and toxic materials). Fractions from CH₂Cl₂ extracts were more effective than fractions from EtOAc extracts. Five herbs (Curcuma wenyujin, Chrysanthemum indicum, Salvia chinensis, Ligusticum chuanxiong Hort. and Cassia tora L.) could sensitise these resistant cancer cells at a non-toxic concentration (10 µg mL⁻¹), and markedly increased doxorubicin accumulation in MCF-7/ADR cells, which necessitates further investigations on the active ingredients of these herbs and their underlying mechanisms.

Topics: Acetates; Cassia; Cell Line, Tumor; Chrysanthemum; Curcuma; Doxorubicin; Drug Resistance, Multiple; Drugs, Chinese Herbal; Humans; Ligusticum; Medicine, Chinese Traditional; Methylene Chloride; Plant Extracts; Plants, Medicinal; Salvia; Tetrazolium Salts; Thiazoles

Monocyte chemoattractant protein-1 (MCP-1) is a potent mediator of macrophage migration and therefore, plays an essential role in early events of inflammation. In the present study, we show the protein kinase C activator, phorbol myristate acetate (PMA), potently induced mRNA expression and secretion of the C-C chemokine MCP-1 in U937 cells. We found that curcumin, a natural biologically active compound extracted from rhizomes of Curcuma species, significantly inhibited the PMA-induced increase in MCP-1 expression and secretion. These effects of curcumin are dose dependent and correlate with the suppression of MCP-1 mRNA expression levels. Curcumin inhibited PMA-mediated activation of extracellular signal-regulated kinase (ERK) and NF-kappaB transcriptional activity. Therefore, one possible anti-inflammatory mechanism of curcumin may be to inhibit the secretions of inflammatory MCP-1 chemokine.

Topics: Antineoplastic Agents; Blotting, Western; Cell Nucleus; Chemokine CCL2; Coloring Agents; Curcumin; Electrophoretic Mobility Shift Assay; Enzyme-Linked Immunosorbent Assay; Extracellular Signal-Regulated MAP Kinases; Humans; Luciferases; NF-kappa B; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA; Tetradecanoylphorbol Acetate; Tetrazolium Salts; Thiazoles; Transcription Factor AP-1; Transcription, Genetic; Transfection; U937 Cells

A variety of cancers ectopically express human chorionic gonadotropin beta (hCGbeta). Patients harboring such cancers have poor prognosis and adverse survival. A recombinant chimeric antibody, cPiPP, exhibiting high affinity and specificity for hCGbeta/hCG was engineered. This study was designed to determine whether this antibody alone or conjugated to curcumin can selectively kill tumor cells expressing hCGbeta.. The study was carried out on MOLT-4 and U-937 cells expressing hCGbeta and on peripheral blood leukocytes of acute myeloid leukemia (AML) patients. The anticancerous compound curcumin was conjugated to cPiPP. The binding of cPiPP and cPiPP-curcumin conjugate to cells was studied by flow cytometry and cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), FACS with propidium iodide staining, trypan blue exclusion assay and microscopy.. The antibody did not impair the growth of MOLT-4 and U-937 cells in culture. Its conjugate with curcumin, however, was lethal to both cell lines. The immunoconjugate killed tumor cells bearing the CD33 marker of an AML patient expressing hCGbeta but did not have a similar action on cells of another AML patient with the CD13 marker but who was negative for hCGbeta.. A humanized antibody against hCGbeta linked to curcumin has potential for therapy of hCGbeta-expressing tumors.

Topics: Aged; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cell Separation; Chorionic Gonadotropin, beta Subunit, Human; Curcumin; Drug Design; Female; Humans; Leukemia; Leukemia, Myeloid, Acute; Leukocytes, Mononuclear; Lymphoma; Male; Middle Aged; Sialic Acid Binding Ig-like Lectin 3; Tetrazolium Salts; Thiazoles; U937 Cells

Curcumin, the active compound of the rhizome of Curcuma longa has anti-inflammatory, antioxidant and antiproliferative activities. This agent has been shown to regulate numerous transcription factors, cytokines, protein kinases, adhesion molecules, redox status and enzymes that have been linked to inflammation. While curcumin has been identified as an activator of apoptosis in several cell lines, the mechanism by which it initiates apoptosis, however, remains poorly understood. We considered curcumin from the point of view of its ability to protect against oxidative stress, the latter being one factor strongly implicated in the development of Parkinson's disease. Although the etiology of Parkinson's disease remains unknown, epidemiological studies have linked exposure to pesticides such paraquat to an increased risk of developing the condition. Analysis of the neurotoxic properties of these pesticide compounds has been focused on their ability to induce oxidative stress in neural cells. Given curcumin's capacity to protect against oxidative stress, it has been considered as a potential therapeutic agent for neurodegenerative diseases such as Parkinson's disease that involve an oxidative stress component. In the present report we describe the effect of curcumin in paraquat-mediated apoptosis of N27 mesencepahlic cells. We show that subtoxic concentrations of curcumin sensitize N27 mesencephalic cells to paraquat-mediated apoptosis.

Topics: Acetylcysteine; Animals; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Caspase 3; Cell Line, Transformed; Curcumin; Dose-Response Relationship, Drug; Drug Synergism; Flow Cytometry; Gene Expression Regulation; Herbicides; Hydrogen Peroxide; Mesencephalon; Neurons; Nitric Oxide Synthase Type I; Nitric Oxide Synthase Type II; Paraquat; Rats; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles; Tyrosine; Vitamin E

Iron overload can cause liver toxicity and increase the risk of liver failure or hepatocellular carcinoma in humans. Curcumin (diferuloylmethane), a component of the food spice turmeric, has antioxidant, iron binding and hepatoprotective properties. The aim of this study was to quantify its effects on iron overload and the resulting downstream toxic effects in cultured T51B rat liver epithelial cells.. T51B cells were loaded with ferric ammonium citrate (FAC) with or without the iron delivery agent 8-hydroxyquinoline. Cytotoxicity was measured by methylthiazolyldiphenyl-tetrazolium bromide assay. Iron uptake and iron bioavailability were documented by chemical assay, quench of calcein fluorescence and ferritin induction. Reactive oxygen species (ROS) were measured by a fluorescence assay using 2',7'-dichlorodihydrofluorescein diacetate. Oxidative stress signalling to jnk, c-jun and p38 was measured by a Western blot with phospho-specific antibodies.. Curcumin bound iron, but did not block iron uptake or bioavailability in T51B cells given FAC. However, it reduced cytotoxicity, blocked the generation of ROS and eliminated signalling to cellular stress pathways caused by iron. Inhibition was observed over a wide range of FAC concentrations (50-500 microM), with an apparent IC(50) in all cases between 5 and 10 microM curcumin. In contrast, desferoxamine blocked both iron uptake and toxic effects of iron at concentrations that depended on the FAC concentration. The effects of curcumin also differed from those of alpha-tocopherol, which did not bind iron and was less effective at blocking iron-stimulated ROS generation.. Curcumin reduced iron-dependent oxidative stress and iron toxicity in T51B cells without blocking iron uptake.

Topics: Animals; Cells, Cultured; Curcumin; Deferoxamine; Epithelial Cells; Ferric Compounds; Inhibitory Concentration 50; Liver; Oxidative Stress; Quaternary Ammonium Compounds; Rats; Reactive Oxygen Species; Signal Transduction; Tetrazolium Salts; Thiazoles

Amyloid beta peptide (Abeta), generated by proteolytic cleavage of the amyloid precursor protein (APP), play a pivotal role in the pathogenesis of Alzheimer's disease (AD). The key step in the generation of Abeta is cleavage of APP by beta-site APP-cleaving enzyme 1 (BACE1). There is increasing evidence supporting an interaction between APP, Abeta and metal ions. Both APP and Abeta affect ion homeostasis. Conversely, metal ions may interact with several AD-associated pathways involved in neurofibrillary tangle formation, secretase cleavage of APP, proteolytic degradation of Abeta and the generation of reactive oxygen species. However, the underlying mechanisms remain elusive. Here we first reported the differential effects of AD-related metal ions at subtoxic concentrations on the transcription levels of APP and BACE1 in PC12 cells. Copper (Cu(2+), 50-100 microM) and manganese (Mn(2+), 50-100 microM) potently increased the expression of both APP and BACE1 in a time- and concentration-dependent pattern, while zinc (Zn(2+)), iron (Fe(2+)) and aluminum (Al(3+)) did not. To uncover the mechanism(s) of the increasing expression by these ions, we observed the effects of several antioxidants and some specific inhibitors on the up-expression of APP and BACE1 by metal ions. Curcumin almost completely blocked the effects of these irons, while minocycline and sodium ferulate slightly suppressed the increased BACE1 mRNA level. Signaling pathway specific inhibitors PD98059, SB203580 and CEP11004 modestly blocked the up-transcription of APP induced by copper. These results suggest that these irons cause differential effects on the expression of APP and BACE1 in PC12 cells, and curcumin can significantly reverse their effects.

Topics: Amyloid beta-Protein Precursor; Amyloid Precursor Protein Secretases; Analysis of Variance; Animals; Curcumin; Dose-Response Relationship, Drug; Enzyme Inhibitors; Gene Expression Regulation; Metals; PC12 Cells; Rats; RNA, Messenger; Tetrazolium Salts; Thiazoles; Time Factors

AMP-activated protein kinase (AMPK), an evolutionarily conserved serine/threonine protein kinase, serves as an energy sensor in all eukaryotic cells. Recent findings suggest that AMPK activation strongly suppresses cell proliferation and induces cell apoptosis in a variety of cancer cells. Our study demonstrated that chemopreventive agent curcumin strongly activates AMPK in a p38-dependent manner in CaOV3 ovarian cancer cells. Pretreatment of cells with compound C (AMPK inhibitor) and SB203580 (p38 inhibitor) attenuates curcumin-induced cell death. We also observed that curcumin induces p53 phosphorylation (Ser 15) and both compound C and SB203580 pretreatment inhibit p53 phosphorylation. Collectively, our data suggest that AMPK is a new molecular target of curcumin and AMPK activation partially contributes to the cytotoxic effect of curcumin in ovarian cancer cells.

Topics: AMP-Activated Protein Kinases; Antineoplastic Agents; Cell Death; Cell Line, Tumor; Cell Survival; Curcumin; Enzyme Inhibitors; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Ovarian Neoplasms; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Tetrazolium Salts; Thiazoles

Leukemias are common worldwide. Wilms' tumor1 (WT1) protein is highly expressed in leukemic blast cells of myeloid and lymphoid origin. Thus, WT1 mRNA serves as a tumor marker for leukemias detection and monitoring disease progression. Curcumin is well known for its anti-cancer property. The objective of this study was to investigate the effect of curcumin on WT1 gene expression in patient leukemic cells. The leukemic cells were collected from 70 childhood leukemia patients admitted at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, in the period July 2003 to February 2005. There were 58 cases of acute lymphoblastic leukemia (ALL), 10 cases of acute myeloblastic leukemia (AML), and 2 cases of chronic myelocytic leukemia (CML). There were 41 males and 29 females ranging from 1 to 15 years old. Leukemic cells were cultured in the presence or absence of 10 mM curcumin for 48 h. WT1 mRNA levels were determined by RT-PCR. The result showed that curcumin reduced WT1 gene expression in the cells from 35 patients (50%). It affected the WT1 gene expression in 4 of 8 relapsed cases (50%), 12 of 24 cases of drug maintenance (50%), 7 of 16 cases of completed treatment (44%), and 12 of 22 cases of new patients (54%). The basal expression levels of WT1 gene in leukemic patient cells as compared to that of K562 cells were classified as low level (1-20%) in 6 of 20 cases (30%), medium level (21-60%) in 12 of 21 cases (57%), and high level (61-100%) in 17 of 23 cases (74%). In summary, curcumin decreased WT1 mRNA in patient leukemic cells. Thus, curcumin treatment may provide a lead for clinical treatment in leukemic patients in the future.

Topics: Adolescent; Antineoplastic Agents; Cell Line, Tumor; Child; Child, Preschool; Curcumin; Electrophoresis, Polyacrylamide Gel; Female; Gene Expression Regulation, Neoplastic; Genes, Wilms Tumor; Humans; Infant; K562 Cells; Leukemia; Male; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tetrazolium Salts; Thiazoles

Curcumin, the yellow pigment in the spice turmeric, has potent chemopreventive activities that involve diverse molecular pathways. It is widely believed that curcumin pro-apoptotic properties are mediated by downregulation of NF kappa B (NFkappaB). The p65/RelA subunit of NFkappaB may influence cell death, in part by activation of NFkappaB anti-apoptotic target genes including X-linked inhibitor of apoptosis (XIAP), A20, bcl-xL and inhibition of sustained activation of c-Jun N-terminal kinase (JNK). We have shown previously that curcumin inhibits NFkappaB, activates JNK and promotes apoptosis in HCT116 colorectal cancer cells. Here, we show that forced overexpression of p65 does not affect curcumin-induced JNK activation. Indeed, overexpression of p65 enhanced curcumin-mediated apoptosis as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay and poly(ADP-ribose) polymerase (PARP) cleavage. This potentiating effect of p65 upon curcumin-mediated apoptosis was reversed by transfection of cells with an IkappaB super-repressor (DeltaNIkappaB). Curcumin treatment inhibited expression of NFkappaB anti-apoptotic target genes in mock-transfected and in p65-overexpressing HCT116 cells, although expression levels remained higher in the latter. Taken together, these results show that curcumin-mediated activation of JNK or induction of apoptosis does not require inhibition of p65. Furthermore, curcumin/p65 synergy in promotion of apoptosis cannot be attributed to active repression of NFkappaB anti-apoptotic genes.

Topics: Apoptosis; Cell Line, Tumor; Cell Survival; Curcumin; Gene Expression Regulation, Neoplastic; Humans; NF-kappa B; Poly(ADP-ribose) Polymerases; Tetrazolium Salts; Thiazoles; Transcription Factor RelA; Transfection; X-Linked Inhibitor of Apoptosis Protein

Inhibition of the COP9 signalosome (CSN) associated kinases CK2 and PKD by curcumin causes stabilization of the tumor suppressor p53. It has been shown that curcumin induces tumor cell death and apoptosis. Curcumin and emodin block the CSN-directed c-Jun signaling pathway, which results in diminished c-Jun steady state levels in HeLa cells. The aim of this work was to search for new CSN kinase inhibitors analogue to curcumin and emodin by means of an in silico screening method.. Here we present a novel method to identify efficient inhibitors of CSN-associated kinases. Using curcumin and emodin as lead structures an in silico screening with our in-house database containing more than 10(6) structures was carried out. Thirty-five compounds were identified and further evaluated by the Lipinski's rule-of-five. Two groups of compounds can be clearly discriminated according to their structures: the curcumin-group and the emodin-group. The compounds were evaluated in in vitro kinase assays and in cell culture experiments.. The data revealed 3 compounds of the curcumin-group (e.g. piceatannol) and 4 of the emodin-group (e.g. anthrachinone) as potent inhibitors of CSN-associated kinases. Identified agents increased p53 levels and induced apoptosis in tumor cells as determined by annexin V-FITC binding, DNA fragmentation and caspase activity assays.. Our data demonstrate that the new in silico screening method is highly efficient for identifying potential anti-tumor drugs.

Topics: Annexin A5; Antineoplastic Agents; Apoptosis; Caspase 3; Caspase 7; Caspases; Cell Line, Tumor; Cell Survival; COP9 Signalosome Complex; Curcumin; DNA Fragmentation; Drug Design; Drug Screening Assays, Antitumor; Emodin; Enzyme Inhibitors; Gene Expression Regulation, Neoplastic; HeLa Cells; Humans; Models, Chemical; Models, Statistical; Multiprotein Complexes; Peptide Hydrolases; Propidium; Proto-Oncogene Proteins c-jun; Signal Transduction; Software; Tetrazolium Salts; Thiazoles; Tumor Suppressor Protein p53

The purpose of this study was to determine whether curcumin would trigger cell death in the head and neck squamous cell carcinoma (HNSCC) cell lines CCL 23, CAL 27, and UM-SCC1 in a dose-dependent fashion.. HNSCC cells were treated with curcumin and assayed for in vitro growth suppression using 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyl tetrazolium bromide and fluorescence-activated cell sorting analyses. Expression of p16, cyclin D1, phospho-Ikappabeta, and nuclear factor-kappabeta (NF-kappabeta) were measured by Western blotting, gel shift, and immunofluorescence.. Addition of curcumin resulted in a dose-dependent growth inhibition of all three cell lines. Curcumin treatment resulted in reduced nuclear expression of NF-kappabeta. This effect on NF-kappabeta was further reflected in the decreased expression of phospho-Ikappabeta-alpha. Whereas the expression of cyclin D1, an NF-kappabeta-activated protein, was also reduced, there was no difference in the expression of p16 at the initial times after curcumin treatment. In vivo growth studies were done using nude mice xenograft tumors. Curcumin was applied as a noninvasive topical paste to the tumors and inhibition of tumor growth was observed in xenografts from the CAL27 cell line.. Curcumin treatment resulted in suppression of HNSCC growth both in vitro and in vivo. Our data support further investigation into the potential use for curcumin as an adjuvant or chemopreventive agent in head and neck cancer.

Topics: Animals; Annexin A5; Antineoplastic Agents; Blotting, Western; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Nucleus; Cell Separation; Cell Survival; Curcumin; Cyclin D1; Dose-Response Relationship, Drug; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; I-kappa B Proteins; In Vitro Techniques; Mice; Mice, Nude; Microscopy, Fluorescence; Neoplasm Transplantation; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Tetrazolium Salts; Thiazoles

While P-glycoprotein (Pgp) and multidrug resistance-associated protein 1 (MRP1) are known to be important in acquired doxorubicin resistance, the role of glutathione S-transferases (GST) remains unclear. Our study assessed roles of these 3 factors in a human drug-sensitive carcinoma cell line (HEp2), a subclone made resistant by prolonged incubation in doxorubicin (HEp2A), and HEp2 cells stably transfected with human GSTP1. Drug-resistant HEp2A cells showed greater total GST activity, GSTP class enzyme expression, Pgp expression, MRP1 transcript expression, drug efflux and at least 13-fold greater resistance to doxorubicin than the parent HEp2 cell line. GSTM class enzyme expression was similar in both cell types, while GSTA class enzymes were not detected. In the resistant HEp2A cells, cytotoxicity was markedly enhanced by the Pgp/MRP inhibitor verapamil at low doxorubicin concentrations. The GST inhibitor curcumin also enhanced cytotoxicity in HEp2A cells when the Pgp/MRP efflux barrier had been reversed by verapamil or overcome by high doxorubicin concentrations. In addition, curcumin had a chemosensitising effect at low doxorubicin concentrations in HEp2 cells. Stable transfection of HEp2 cells with human GSTP1 increases doxorubicin resistance 3-fold over control cells. Our study indicates involvement of GSTP enzymes as well as efflux mechanisms in the acquired doxorubicin-resistance phenotype.

Topics: Animals; Antineoplastic Agents; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP-Binding Cassette Transporters; Calcium Channel Blockers; Coloring Agents; Curcumin; Cytosol; Dose-Response Relationship, Drug; Doxorubicin; Drug Resistance, Neoplasm; Genetic Vectors; Glutathione S-Transferase pi; Glutathione Transferase; Humans; Immunoblotting; Inhibitory Concentration 50; Isoenzymes; Liver; Multidrug Resistance-Associated Proteins; Phenotype; Protein Transport; Rats; Reverse Transcriptase Polymerase Chain Reaction; Tetrazolium Salts; Thiazoles; Transfection; Tumor Cells, Cultured; Verapamil

Curcumin, a dietary pigment in turmeric, posseses anti-carcinogenic and anti-metastatic properties. The present study was conducted to study in vitro chemopreventive effects of curcumin in transformed breast cells. Here, we show that curcumin inhibits H-ras-induced invasive phenotype in MCF10A human breast epithelial cells (H-ras MCF10A) and downregulates matrix metalloproteinase (MMP)-2 dose-dependently. Curcumin exerted cytotoxic effect on H-ras MCF10A cells in a concentration-dependent manner. Curcumin-induced cell death was mainly due to apoptosis in which a prominent downregulation of Bcl-2 and upregulation of Bax were involved. We also suggest a possible involvement of caspase-3 in curcumin-induced apoptosis. Curcumin treatment resulted in the production of reactive oxygen species (ROS) in H-ras MCF10A cells. Apoptotic event by curcumin was significantly inhibited by pretreatment of an antioxidant N-acetyl-L-cysteine (NAC), suggesting redox signaling as a mechanism responsible for curcumin-induced apoptosis in H-ras MCF10A cells. Taken together, our results demonstrate that curcumin inhibits invasion and induces apoptosis, proving the chemopreventive potential of curcumin.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Breast; Caspase 3; Caspases; Curcumin; DNA Fragmentation; Enzyme Activation; Epithelial Cells; Female; Gene Expression Regulation; Genes, bcl-2; Genes, ras; Humans; Immunoblotting; Oxidation-Reduction; Reactive Oxygen Species; Signal Transduction; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured

Pharmacologically safe compounds that can inhibit the proliferation of tumor cells have potential as anticancer agents. Curcumin, a diferuloylmethane, is a major active component of the food flavor turmeric (Curcuma longa) that exhibits anticarcinogenic properties in vivo. In vitro, it suppressed c-jun/Ap-1 and NF-kappaB activation and type 1 human immunodeficiency virus long-terminal repeat-directed gene expression. We examined the antiproliferative effects of curcumin against several breast tumor cell lines, including hormone-dependent and -independent and multidrug-resistant (MDR) lines. Cell growth inhibition was monitored by [3H]thymidine incorporation, Trypan blue exclusion, crystal violet dye uptake and flow cytometry. All the cell lines tested, including the MDR-positive ones, were highly sensitive to curcumin. The growth inhibitory effect of curcumin was time- and dose-dependent, and correlated with its inhibition of ornithine decarboxylase activity. Curcumin preferentially arrested cells in the G2/S phase of the cell cycle. Curcumin-induced cell death was neither due to apoptosis nor to any significant change in the expression of apoptosis-related genes, including Bcl-2, p53, cyclin B and transglutaminase. Overall our results suggest that curcumin is a potent antiproliferative agent for breast tumor cells and may have potential as an anticancer agent.

Topics: Antibiotics, Antineoplastic; Antineoplastic Agents; Blotting, Western; Breast Neoplasms; Cell Cycle; Cell Division; Cell Line; Curcumin; Doxorubicin; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Humans; Ornithine Decarboxylase; Tetrazolium Salts; Thiazoles