Caucasian Colon Adenocarcinoma Research Articles

Synthesis and Evaluation of 6‐Amino‐2‐styrylchromones as Anticancer Agents

Abstract2‐Styrylchromones, a C‐2 styrene derivative of chromones, possess a broad spectrum of biological activity. However, 2‐styrylchromones are scarce in nature but not to mention the amino‐substituted styrylchromones. We have synthesized fourteen 6‐amino‐2‐styrylchromones and evaluated their anti‐cancer activity on human caucasian colon adenocarcinoma cells (HT‐29). 6‐Amino‐2‐styrylchromones and related derivatives have never been revealed their pharmacology profiles. Among the novel fourteen compounds, we reported that compound 9 h exhibited its potential with the comparable activity with 5‐flurouracil (5‐FU) as a referenced drug.

Targeting cellular gaps using Janus nanoparticles containing cationic polymers and surfactant lipids.

Since nanoparticles are taken up into cells by endocytosis, phagocytosis, or pinocytosis, they have been studied as intracellular drug carriers. Janus particles have an anisotropic structure composed of two or more distinct domains and have been proposed for use in various applications, including use as imaging agents or nanosensors. This study aimed to clarify the influence of the type of nanoparticles on their distribution in a human Caucasian colon adenocarcinoma (Caco-2) cell monolayer. We fabricated Janus and conventional spherical nanoparticles composed of pharmaceutically applicable ingredients. Janus and spherical nanoparticles composed of a cationic polymer and surfactant lipids were prepared by controlling the solvent removal pattern from the oil phase in the solvent removal process using the solvent evaporation and solvent diffusion methods. The distribution of nanoparticles in the Caco-2 cell monolayer was then evaluated using confocal laser microscopy. The mean hydrodynamic size of the fabricated Janus nanoparticles was 119.2 ± 4.6 nm. Distribution analysis using Caco-2 cells suggested that Janus nanoparticles were localized around the adherens junctions located just below the tight junction. Clear localization was not observed in non-Janus nanoparticles with the same composition. The clear localization of the Janus nanoparticles around the adherens junction may be due to their positive charge and asymmetric structure. Our results suggest the considerable potential for the development of nanoparticulate drug carriers to target cellular gaps.

Tracking the Evolution of Metastasis with Self-Functionalized 3D Nanoprobes.

Despite recent advances in cancer treatment, metastasis is the cause of mortality in 90% of cancer cases. It has now been well-established that dissemination of cancer cells to distant sites occurs very early during tumorigenesis, resulting in the minimal effect of surgical or chemotherapeutic treatments after the detection of metastasis. The underlying reason for this challenge is mostly due to the limited understanding of molecular mechanisms of the metastasis cascade, particularly related to metastatic traits. Therefore, there is an urgent need to investigate this currently invisible evolution of metastasis. The tracking of metastasis evolution has not been addressed yet. Here, we introduce, for the first time, a synchronous approach to unveil the molecular mechanisms of the metastasis cascade. As cancer stem cells (CSCs) demonstrate cancer initiation, drug resistance, metastasis, and tumor relapse and can exist in a quasi-intermediate epithelial-mesenchymal transition state, the tumor-initiating events during a CSCs metamorphosis were monitored with single-cell sensitivity. Because of the invasive and resistive properties of the metastable intermediate CSCs, investigation of the molecular profiles of the quasi-intermediate CSCs was necessary for the detection of metastasis dissemination. For this purpose, the ultrasensitive technique of surface-enhanced Raman scattering (SERS) was adopted. Titanium-based, biocompatible three-dimensional (3D) nanoprobes that were synthesized for multiphoton ionization achieved a substantial SERS enhancement of ∼80-fold due to the oxygen vacancy-enriched composition of the nanoprobes. The 3D interconnected complex nanoarchitecture of the nanoprobes enabled us to entrap the nonadherent CSCs of three metastatic cancer cell lines (triple negative breast adenocarcinoma (MDAMB231), human Caucasian colon adenocarcinoma (COLO 205), and cervical adenocarcinoma (HeLa)─all very aggressive forms of cancer). The nanoprobes not only promoted the CSC proliferation to successfully attain the quasi-intermediate states but also monitored its reprogramming into a cancer cell state. The nanoprobes substantially amplified weak intracellular Raman signals to capture the molecular events during a CSC transformation. The detection of cancer was achieved with 100% accuracy. We experimentally demonstrated that the molecular signatures of CSC reprogramming are cancer-type specific. This observation enabled us to identify the origin of metastasis with 100% accuracy, providing more clarity on the relatively unknown quasi-intermediate states. This first demonstration of CSC-based tracking of metastasis evolution has the potential to provide an insightful perspective of tumorigenesis that could be useful in cancer diagnosis and prognosis as well as in the monitoring of therapeutic interventions.

Prognostic significance of Notch3 immunoreactivity patterns in Caucasian colon adenocarcinoma patients

IntroductionColon cancer is traditionally regarded as the most commonly diagnosed gastrointestinal malignant disease. Nevertheless, despite advances in diagnosis and novel therapeutic options, the clinical outcomes of patients are still not satisfactory.AimTo investigate the prognostic role of Notch3, an immunohistochemical study was performed on samples of colon adenocarcinoma tissues and adjacent non-pathological mucosa in Caucasian patients.Material and methodsParaffin-embedded colon adenocarcinoma samples were assessed immunohistochemically for Notch3 protein. Connections between Notch3 immunoexpression and clinicopathological factors including the 5-year overall survival (OS) were evaluated.ResultsThe level of the Notch3 immunohistochemical reactivity was correlated with the grade of the histological differentiation (p < 0.001). A strong expression of Notch3 protein was detected more frequently in patients with G3 tumours than in patients with G1 tumours. Moreover, the significant difference was also detected between the patients with G1 tumour and those with G2. In the G2 patients a strong reactivity was described more frequently. The Kaplan-Meier survival analysis showed that the overall survival rate in the group of patients with a low expression level of Notch3 was significantly greater than that for patients with a moderate or strong level of Notch3 immunoreactivity (p < 0.001). A multivariate analysis demonstrated that the grade of tumour differentiation (p = 0.001) and Notch3 expression (p < 0.001) were independent risk factors for worse survival.ConclusionsThe high level of Notch3 immunoexpression was demonstrated to be associated with malignancy-related clinicopathological factors and 5-year overall survival of patients.

Low doses of imidacloprid induce disruption of intercellular adhesion and initiate proinflammatory changes in Caco-2 cells

Imidacloprid is the most widely used pesticide of the neonicotinoid class. Neonicotinoid toxicities against various insects are well known. Nevertheless, there are rising evidences that neonicotinoids exert cytotoxic effects on different non-target organisms including mammals, fish, birds etc. Besides, depending on pesticide application, the exposed plants absorb some part of used neonicotinoids and their residues are detected in agricultural products worldwide. Thus, the continuous consumption of fruits and vegetables contaminated with neonicotinoids is a high risk factor for humans despite the low doses. Intestine epithelial cells are the first targets of the neonicotinoid cytotoxicity in humans because of its direct way of administration. The epithelial cells provide the barrier function of the intestinal system via specialized intercellular adhesion. The effects of imidacloprid on the intestine barrier function and inflammatory cytokines production are still unknown. In the present study, we exposed the human Caucasian colon adenocarcinoma (Caco-2) epithelial cells to low doses (0.10–0.75 µg/mL) of imidacloprid in order to assess the expression of tight and adherens junctions proteins, occludin and E-cadherin, and production of proinflammatory cytokine TNF α and iNOS. Imidacloprid induced dose-dependent decline in both occludin and E-cadherin levels. By contrast, TNF-α and iNOS contents were upregulated in imidacloprid-exposed Caco-2 cells. Decrease in tight and adherens junctions proteins indicates that the barrier function of intestine epithelial cells could be damaged by imidacloprid administration. In addition, TNF-α and iNOS upregulation indicates that imidacloprid is potent to activate proinflammatory response in enterocytes. Thus, imidacloprid can affect intestine barrier function through the increase of proinflammatory cytokine production and decrease in adhesiveness of enterocytes. The further assessment of the role of adhesion proteins and inflammatory cytokines in neonicotinoid pesticide cytotoxicity as it affects enterocyte barrier function is required to highlight the risk factor of use of neonicotinoids.

Roles and mechanisms of circulating CEACAM1 in the cirrhosis-related intestinal hyperpermeability: in vitro approach.

Cirrhosis-related intestinal hyperpermeability and endotoxemia are characterized by intestinal epithelial cell apoptosis, impaired restitution (proliferation and migration), decreased tight junction protein levels, and subsequent barrier dysfunction. In addition to endotoxin and tumor necrosis factor-α (TNFα), carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) plays crucial roles in the regulation of apoptosis, restitution, tight junction protein-maintained barrier function of intestinal epithelial cells. This study aims to explore the roles and underlying mechanisms of CEACAM1 in cirrhosis-related intestinal hyperpermeability through in vitro approach. In cirrhotic patients, high serum levels of intestinal hyperpermeability (zonulin and endotoxin) markers were accompanied by elevated serum levels of TNFα and soluble CEACAM1. In in vitro experiments, we evaluated the individual and interacted roles of TNFα and human recombinant CEACAM1 (hrCEACAM1) in LC-sera (sera of cirrhotic patients)-induced intestinal hyperpermeability-related pathogenic signals. In the cell Line human from human colon (Caucasian colon adenocarcinoma) (Caco-2) cell culture, LC-sera, TNFα, and hrCEACAM1 increased apoptosis (measured by Terminal deoxynucleotidyl transferase [TdT] dUTP nick end labeling+/annexin-5+propidium iodide+ cells and caspase-3 activity), decreased restitution capacity (proliferation and migration), and disrupted tight junction protein-maintained barrier function in Caco-2 cells. The pathogenic changes mentioned above were accompanied by an increase in intracellular reactive oxygen species (ROS) levels, lactate dehydrogenase release, and endoplasmic reticulum stress-related signals in the LC-sera or TNFα-pretreated Caco-2 cells. Concomitant incubation of Caco-2 cells with anti-CEACAM1 suppressed these LC-sera or TNFα-induced negative effects on restitution, barrier function, and cell viability. This study demonstrated that sera from cirrhotic patients contain soluble CEACAM1, which is involved in the pathogenesis of intestinal hyperpermeability. Accordingly, it is noteworthy to explore the potential use of anti-CEACAM1 treatment for cirrhosis-related intestinal hyperpermeability and endotoxemia.

Gellan Gum Promotes the Differentiation of Enterocytes from Human Induced Pluripotent Stem Cells.

The evaluation of drug pharmacokinetics in the small intestine is critical for developing orally administered drugs. Caucasian colon adenocarcinoma (Caco-2) cells are employed to evaluate drug absorption in preclinical trials of drug development. However, the pharmacokinetic characteristics of Caco-2 cells are different from those of the normal human small intestine. Besides this, it is almost impossible to obtain primary human intestinal epithelial cells of the same batch. Therefore, human iPS cell-derived enterocytes (hiPSEs) with pharmacokinetic functions similar to human intestinal epithelial cells are expected to be useful for the evaluation of drug absorption. Previous studies have been limited to the use of cytokines and small molecules to generate hiPSEs. Dietary fibers play a critical role in maintaining intestinal physiology. We used gellan gum (GG), a soluble dietary fiber, to optimize hiPSE differentiation. hiPSEs cocultured with GG had significantly higher expression of small intestine- and pharmacokinetics-related genes and proteins. The activities of drug-metabolizing enzymes, such as cytochrome P450 2C19, and peptide transporter 1 were significantly increased in the GG treatment group compared to the control group. At the end point of differentiation, the percentage of senescent cells increased. Therefore, GG could improve the differentiation efficiency of human iPS cells to enterocytes and increase intestinal maturation by extending the life span of hiPSEs.

A novel strategy for screening bioavailable quality markers of traditional Chinese medicine by integrating intestinal absorption and network pharmacology: Application to Wu Ji Bai Feng Pill

BackgroundMajor components are often used as marker compounds for quality control of traditional Chinese medicines (TCMs). However, these compounds may not necessarily bioavailable and active in vivo, thereby, failing to control the “quality”. PurposeThe purpose of this paper is to develop a novel strategy integrating absorption and activity deduced from network pharmacology to identify more reasonable markers for quality control of TCM formulas using Wu Ji Bai Feng Pill (WJBFP) as an example. Study DesignHuman Caucasian colon adenocarcinoma (Caco-2) cell transport studies and a bioavailability-enhanced network pharmacological approach were integrated to identify better phytochemical markers for quality control. MethodsThe absorption of multiple components in WJBFP was evaluated by a Caco-2 cell culture model. Nine databases were used to identify potential targets in the network pharmacology analysis. Cytoscape 3.7.2 was employed for the network data integration, visualization, and centrality analysis. Molecular docking was carried out to investigate the binding affinity of the identified markers to their candidate targets. ResultsThe apparent permeability coefficient (Papp) and efflux ratio (ER) of 66 compounds were determined. Five hundred and two putative targets and 187 primary dysmenorrhea (PD) related targets were identified. Twenty-two candidate targets interacting with 20 potential active compounds were screened with the putative PD related targets intersection network using Degree Centrality (DC) ranking. By integrating absorption, 16 candidate targets interacting with 8 potential active compounds were identified. Besides, 53 compounds hitting candidate targets were divided into two classes according to their DC values. Then each of the two classes of DC was stratified into two groups based on the Papp for a total of four classes. Finally, five compounds belonging to Class 1 with higher DC and higher Papp, formononetin, ferulic acid, isoliquiritigenin, neocryptotanshinone and senkyunolide A, were identified as potential bioavailable phytochemical markers for the quality control of WJBFP against PD. Furthermore, molecular docking analysis validated the interplay between candidate targets and marker ingredients. ConclusionA novel strategy combining intestinal absorption with network pharmacology analysis was successfully established to identify bioavailable and bioactive markers for quality control of WJBFP against PD.

Study of the intestinal uptake and permeability of gold nanoparticles using both in vitro and in vivo approaches

Gold nanoparticles (AuNPs) are highly attractive to biomedical applications. Here, we investigated the effects of (i) ca. 15 nm spherical AuNPs capped with citrate or 11-mercaptoundecanoic acid (MUA) and (ii) ca. 60 nm spherical citrate-capped AuNPs, and ca. 60 nm MUA-capped star-shaped AuNPs on the cytotoxicity, cellular uptake and permeability, using media supplemented or not with 1% fetal bovine serum (FBS) on caucasian colon adenocarcinoma Caco-2 cells. In addition, the colloidal stability of the nanoparticles in media (supplemented or not) was assessed after 24 h-incubations at 60 μM. The 60 nm gold nanospheres and stars were administrated orally to Wistar rats in order to evaluate their systemic absorption and biodistribution after 24 h. At non-supplemented media settings, citrate-capped gold nanoparticles seem to be more toxic than their MUA-capped counterparts. Also, smaller nanoparticles show higher toxicity than larger ones. The use of cell culture media with 1% FBS not only increased the stability of all AuNPs, as also significantly reduced their cytotoxicity. In the uptake studies, higher AuNPs incorporation was noticed in serum supplemented media, this effect being particularly significant for the 60 nm nanoparticles. Cellular incorporation depended also on the capping agent and size. None of the tested samples crossed the in vitro intestinal barrier. Confirming the in vitro results, the in vivo biodistribution study of the 60 nm AuNPs orally given to rats showed that their systemic absorption is low and that they are mainly eliminated through the faeces. Altogether, these preliminary results suggest that our novel AuNPs have high potential to be considered promising candidates for application in diagnostics or drug delivery at the intestinal level, showing high biocompatibility. However, unless it is desired that these nanomaterials avoid systemic absorption upon oral administration, additional functionalization should be sought to increase their low bioavailability.

Janus microspheres for enhanced enteral drug delivery: Preparation and orientated attachment to a Caco-2 monolayer.

Conventional oral preparations generally release incorporated drugs omnidirectionally, including into the lumen, leading to a low bioavailability of drugs that are unstable in the gastrointestinal tract. Here, we designed Janus microspheres for efficient mucosal drug delivery as single-sided-release microspheres with the oriented attachment to mucus and evaluated their attachment to and orientation on a Caco-2 (human Caucasian colon adenocarcinoma cell line) monolayer. The microspheres comprised a mucus-oriented hemisphere of an ammonioalkyl methacrylate copolymer and a protective hemisphere of a hard fat. Fluorescein isothiocyanate-dextran with an average molecular weight of 3,000-5,000 Da (FD4) was used as a model hydrophilic drug. A water-in-oil emulsion-type solvent evaporation method was employed for fabrication of the Janus microspheres. The yield of Janus microspheres was found to be dependent on the polymer-to-hard fat ratio, with a maximum yield of over 90% being obtained at a ratio of 1:2, whereas lower and higher ratios resulted in monolithic or star-shaped microspheres. FD4 was specifically localized in the polymeric hemisphere. A cell culture study revealed that the Janus microspheres attached to a Caco-2 monolayer via their polymeric hemispheres with the hard fat hemisphere providing a protective sealing. This may lead to the development of an effective enteral drug delivery system for biomedicines, such as polypeptides and nucleic acids.

No evidence for interactions of dimethylfumarate (DMF) and its main metabolite monomethylfumarate (MMF) with human cytochrome P450 (CYP) enzymes and the P-glycoprotein (P-gp) drug transporter.

Dimethylfumarate (DMF) has long been used as part of a fixed combination of fumaric acid esters (FAE) in some European countries and is now available as an oral monotherapy for psoriasis. The present investigation determined whether DMF and its main metabolite monomethylfumarate (MMF) interact with hepatic cytochrome P450 (CYP) enzymes and the P‐glycoprotein (P‐gp) transporter, and was performed as part of DMF's regulatory commitments. Although referred to in the available product labels/summary of product characteristics, the actual data have not yet been made publicly available. In vitro inhibition experiments using CYP‐selective substrates with human liver microsomes showed 50% inhibitory concentrations (IC50) of >666 µmol/L for DMF and >750 µmol/L for MMF. MMF (≤250 μmol/L; 72 hours) was not cytotoxic in cultured human hepatocyte experiments and mRNA expression data indicated no CYP induction by MMF (1‐250 µmol/L). DMF (≤6.66 mmol/L) showed moderate‐to‐high absorption (apparent permeability [Papp] ≥2.3‐29.7 x 10−6 cm/s) across a Caucasian colon adenocarcinoma (Caco‐2) cell monolayer, while MMF (≤7.38 mmol/L) demonstrated low‐to‐moderate permeability (Papp 1.2‐8.9 × 10−6 cm/s). DMF was not a substrate for P‐gp (net efflux ratios ≤1.22) but was a weak inhibitor of P‐gp at supratherapeutic concentrations (estimated IC50 relative to solvent control of 1.5 mmol/L; [3H]digoxin efflux in Caco‐2 cells). This inhibition is unlikely to be clinically relevant. MMF was not a substrate or inhibitor of P‐gp. Thus, DMF and MMF should not affect the absorption, distribution, metabolism or excretion of coadministered drugs that are CYP and P‐gp substrates.

Validation of an Ex Vivo Permeation Method for the Intestinal Permeability of Different BCS Drugs and Its Correlation with Caco-2 In Vitro Experiments.

The absorption study of drugs through different biological membranes constitutes an essential step in the development of new pharmaceutical dosage forms. Concerning orally administered forms, methods based on monolayer cell culture of Caco-2 (Caucasian colon adenocarcinoma) have been developed to emulate intestinal mucosa in permeability studies. Although it is widely accepted, it has disadvantages, such as high costs or high technical complexity, and limitations related to the simplified structure of the monolayer or the class of molecules that can be permeated according to the transport mechanisms. The aim of this work was to develop a new ex vivo methodology which allows the evaluation of the intestinal apparent permeability coefficient (Papp) while using fewer resources and to assess the correlation with Caco-2. To this end, pig (Sus scrofa) duodenum segments were mounted in Franz diffusion cells and used to permeate four different drugs: ketorolac tromethamine (Kt), melatonin (Mel), hydrochlorothiazide (Htz), and furosemide (Fur). No statistically significant differences (p > 0.05) were observed corelating Papp values from Franz diffusion cells and Caco-2 cell experiments for Kt, Htz, and Fur. However, there were statistically significant differences (p < 0.05) correlating Papp values and Mel. The difference is explained by the role of Mel in the duodenal epithelial paracellular permeability reduction. Ex vivo permeation may be an equivalent method to Caco-2 for drugs that do not produce intestinal membrane phenomena that could affect absorption.

Systematic investigation of in vitro and in vivo safety, toxicity and degradation of mesoporous silica nanoparticles synthesized using commercial sodium silicate

Abstract Mesoporous silica nanoparticles (MSNs) have been extensively investigated as drug delivery carriers due to their unique properties. Although MSNs synthesized using a very economic source of silica, sodium silicate, are promising carriers for drug delivery, information regarding their biocompatibility and in vivo toxicity is scarce. In the present work, biocompatibility, toxicity profile and degradation of mesoporous silica nanoparticles synthesized using commercial sodium silicate have been studied. In vitro degradation studies confirmed that MSNs got degraded to silicic acid within 6 days in PBS 7.4 The synthesized MSNs were found to be biocompatible to different cell lines such as human embryonic kidney (HEK-293) cells, human caucasian colon adenocarcinoma (Caco-2) cells, human liver carcinoma (HepG2) cells and mouse fibroblast (3T3) cells with greater than 90% cell viability at concentration of 200 μg/mL. They did not cause significant hemolysis (only 1.87%) up to 200 μg/mL concentration when incubated for 2 h with chicken red blood cells. Furthermore, hematological and biochemical tests of blood samples as well as histopathological examinations of tissues after MSN injection, confirmed that the MSNs were well tolerated by mice up to 40 mg/kg. Injected MSNs were completely excreted out through urine and feces by rats (injected intravenously with 20 mg/kg of MSNs) within 4 days in form of silicic acid.

Neomacrophorin and premacrophorin congeners from Trichoderma sp. 1212-03

Six novel neomacrophorin and four novel premacrophorin congeners were isolated from the fungus Trichoderma sp. 1212-03. Their planar structures and relative configurations were determined by conventional NMR and mass spectrometry techniques. The electronic circular dichroism (ECD) spectral comparisons with known neomacrophorins were effective for elucidating the structures of congeners carrying 2,3-epoxybenzoquinone (1, 2) and 2,3-epoxybenzosemiquinol substructures (3, 4, 5, 7, 8, and 9). The configuration of the 6-hydroxy-2-cyclohexene-1,4-dione moiety in 5′-deoxyneomacrophorin IV (6) was elucidated by NMR spectral comparison with those of known purpurogemutantidin (16) and mancrophorin F (17). This also resulted in the configurational revision of 16. The relative configuration of the 5,6-epoxy-2-cyclohexen-1,4-diol in premacrophorintriol-II (10) was determined by 13C chemical shift analysis using density functional theory calculations at the ωB97X-D/6-31G* level of theory. Compounds 1 and 6 exhibited potent cytotoxicity against human Caucasian colon adenocarcinoma cell line COLO 201.

Phytoconstituents from Vernonia glaberrima Welw. Ex O. Hoffm. leaves and their cytotoxic activities on a panel of human cancer cell lines

Vernonia glaberrima is a medicinal plant that is used in African traditional medicine for the treatment of skin cancer. The aim of this study was to investigate the anticancer activity of V. glaberrima leaves and isolate its bioactive constituents. Crude methanolic leaves extract of V. glaberrima showing significant cytotoxic activity against cancer cell lines was subjected to chromatographic separation, purification and hydrolysis to yield four compounds namely, nonacosanoic acid, lupeol, 5-methylcoumarin-4-β-glucoside and 4-hydroxy-5-methylcoumarin. Three of the isolated compounds showed significant cytotoxic activity against human malignant melanoma (A375) cell line (IC50: 59.18±2.70 to 139.53±10.79μg/mL) and human caucasian colon adenocarcinoma (HT-29) cell line (IC50: 4.22±0.13 to 20.0±1.91μg/mL) while only lupeol displayed significant activity against human breast adenocarcinoma (MCF7) (IC50: 34.15±2.32μg/mL) cell line. CDK2 receptor and CAIX were identified through molecular docking as potential targets for the bioactive compounds. The findings of this study have revealed the therapeutic potential of V. glaberrima against breast cancer, skin cancer and colorectal carcinoma, respectively and further support its traditional uses in the treatment of skin cancer.

Crude Extracts of Marine-derived and Soil Fungi of the Genus Neosartorya Exhibit Selective Anticancer Activity by Inducing Cell Death in Colon, Breast and Skin Cancer Cell Lines.

Background:The crude ethyl acetate extracts of marine-derived fungi Neosartorya tsunodae KUFC 9213 (E1) and N. laciniosa KUFC 7896 (E2), and soil fungus N. fischeri KUFC 6344 (E3) were evaluated for their in vitro anticancer activities on a panel of seven human cancer cell lines.Materials and Methods:The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed, after 48 h treatments with different concentrations of extracts, to determine their concentration of the extract or Dox that inhibits cell viability by 50% for each cell line. The effects of the crude extracts on DNA damage, clonogenic potential and their ability to induce cell death were also assessed.Results:E1 was found to the void of anti-proliferative effects. E2 was shown to decrease the clonogenic potential in human colorectal carcinoma cell line (HCT116), human malignant melanoma cell line (A375), human breast adenocarcinoma cell line (MCF7), and human caucasian colon adenocarcinoma Grade II cell line (HT29) cells, whereas E3 showed such effect only in HCT116 and MCF7 cells. Both extracts were found to increase DNA damage in some cell lines. E2 was found to induce cell death in HT29, HCT116, MCF7, and A375 cells while extract E3 increased cell death in MCF7 and HCT116 cell lines.Conclusion:The results reveal that E2 and E3 possess anticancer activities in human colon carcinoma, breast adenocarcinoma, and melanoma cells, validating the interest for an identification of molecular targets involved in the anticancer activity.SUMMARY The crude ethyl acetate extract of N. tsunodae (E1) did not decrease cell viability in any of the tested cell linesThe crude ethyl acetate extracts of N. laciniosa (E2) and N. fischeri (E3) decreased cell proliferation in some human cancer cell lines tested at both short- and long-termN. laciniosa (E2) induced a significant increase in the number of cell death, in part, due to the induction of DNA damageN. fischeri (E3) induce cell death but in some cell lines without induction of DNA damage detected by comet assayCrude ethyl extracts of N. laciniosa (E2) and N. fischeri (E3) exert an anticancer activity in human colon carcinoma, breast adenocarcinoma, and malignant melanoma cells. Abbreviations Used: A375: Human malignant melanoma cell line; A549: Human non-small lung cancer cell line; DAPI: 4,6-diamidino-2-phenylindole; DMEM: Dulbecco's Modified Eagle Medium; DMSO: Dimethylsulfoxide; Dox: Doxorubicin; E1: Neosartorya tsunodae KUFC 9213; E2: Neosartorya laciniosa KUFC 7896; E3: Neosartorya fischeri KUFC 6344; FBS: Fetal bovine serum; HCT116: Human colorectal carcinoma cell line; HEPES: (N-[2-hydroxyethyl] piperazine-N’-[2-ethane-sulfonic acid]); HepG2: Human hepatocellular carcinoma cell line; HT29: Human caucasian colon adenocarcinoma Grade II cell line; IC50: Concentration of the extract or Dox that inhibits cell viability by 50%; MCF7: Human breast adenocarcinoma cell line; MEM: Minimum Essential Medium Eagle; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NCI-H460: Human non-small lung cancer cell line; PBS: Phosphate buffered saline; PE: Plating efficiency; RPMI: Roswell park memorial institute medium; SF: Surviving fraction; U-251: Human malignant glioblastoma cell line.

39 Photodynamic therapy of intractable purulent wounds with photoditazine-polymer complex immobilized on hydroxyapatite nanoparticles

Treatment of 5,10,15,20-tetra(pentafluorophenyl) porphyrins with polyethylene glycol (PEG) halides and sodium sulfide is a mild and efficient route to water-soluble PEG substituted porphyrins. The method has been used to access a series of free-base and metallated PEG porphyrins, which have been investigated for photodynamic activity against Human Caucasian colon adenocarcinoma (Caco2) cells. Toxicity, in the presence and absence of light, is shown to be affected by the length of the PEG chain, nature of coordinated metal and axial ligand.

Silver nanoparticles: Antibacterial activity against wound isolates &invitro cytotoxic activity on Human Caucasian colon adenocarcinoma

ObjectiveTo synthesize the silver nanoparticles (AgNPs) using the extracts of Hypnea sp. and to investigate the antibacterial activity against Eshcherichia coli, Staphylococcus aureus and invitro cytotoxic activity on HT-29. MethodsIn the present study, AgNPs were synthesized using the aqueous extract of marine macro-algae, and were characterized using UV-visible spectroscopy, Fourrier Transform Infra red (FT-IR) spectroscopy, X-ray diffraction (XRD) analysis and Transmission Electron Microscopy (TEM) analysis. Further these synthesized AgNPs were evaluated for their antibacterial activity with the clinical isolates from wound specimens. The isolates were characterized by different tests viz., microscopical observation, colony morphology, biochemical & sugar fermentation tests. The synthesized AgNPs were tested for its antibacterial activity against the isolates by agar well diffussion method. The AgNPs were assessesd for its cytotoxic activity on Human Caucasian colon adenocarcinoma (HT-29) cell lines. ResultsIn this study, it is clear that the synthesized AgNPs were spherical measuring 10–20nm and was found to be more bactericidal against Gram-negative bacteria (E. coli) than Gram-positive bacteria (Staphylococcus aureus) isolated from wound specimen. The invitro screening of the AgNPs showed potential cytotoxic activity against the colon cancer cell lines. ConclusionsProteins can bind to nanoparticles either through the electrostatic attraction of negatively charged carboxylate groups in Hypnea sp. and stabilization of the AgNPs by protein occurs. The antimicrobial activities of AgNPs are influenced by the dimensions of the particles the smaller the particles, the greater antimicrobial effect. The cytotoxic activity may be due to the presence of alkaloids present in the Hypnea sp.

Polyacrylamide Nanoparticles as a Delivery System in Photodynamic Therapy

Nanoparticles can be targeted towards, and accumulate in, tumor tissue by the enhanced permeability and retention effect, if sequestration by the reticuloendothelial system (RES) is avoided. The application of nanoparticles in the field of drug delivery is thus an area of great interest, due to their potential for delivering high payloads of drugs site selectively. One area which may prove to be particularly attractive is photodynamic therapy, as the reactive oxygen species (ROS) which cause damage to the tumor tissue are not generated until the drug is activated with light, minimizing generalized toxicity and giving a high degree of spatial control over the clinical effect. In the present study, we have synthesized two types of nanoparticles loaded with photodynamic sensitizers: polylysine bound tetrasulfonato-aluminum phthalocyanine entrapped nanoparticles (PCNP) and polylysine bound tetrasulfonato-aluminum phthalocyanine entrapped nanoparticles coated with a second, porphyrin based, photosensitizer (PCNP-P) to enhance the capacity for ROS generation, and hence therapeutic potential. The mean sizes of these particles were 45 ± 10 nm and 95 ± 10 nm respectively. Uptake of the nanoparticles by human Caucasian colon adenocarcinoma cells (HT29) was determined by flow cytometry and confocal microscopy. Cell viability assays using PCNP-P and PCNP corresponding to the minimum uptake time (<5 min) and maximum uptake time (25 h) demonstrated that these cancer cells can be damaged by light activation of these photodynamic nanoparticles both in the external media and after internalization. The results suggest that, in order to induce photodynamic damage, the nanoparticles need only to be associated with the tumor cell closely enough to deliver singlet oxygen: their internalization within target cells may not be necessary. Clinically, this could be of great importance as it may help to combat the known ability of many cancer cells to actively expel conventional anticancer drugs.

Investigation of Orlistat effects on PXR activation and CYP3A4 expression in primary human hepatocytes and human intestinal LS174T cells

Drugs for weight loss have been in use for nearly hundred years. Orlistat ( Xenical) is a non-centrally acting anti-obesity drug that inactivates gastric and intestinal lipases, thus, preventing absorption of dietary triglycerides. There are reports indicating that Orlistat reduces bioavailability of Cyclosporin to a clinically relevant degree. Since Cyclosporin is metabolized by cytochrome P450 CYP3A4, we examined whether interaction between Orlistat and Cyclosporin involves induction of CYP3A4. Human Caucasian colon adenocarcinoma cells LS174T and primary cultures of human hepatocytes were used, as in vitro models of intestinal and hepatic cells, respectively. Treatment of LS174T cells for 24 h with Orlistat (1–100 mg/L) did not cause induction of CYP3A4 mRNA levels as compared to control cells while Orlistat (100 mg/L) slightly induced CYP3A4 mRNA in human hepatocytes. Rifampicin, a model CYP3A4 inducer, significantly induced CYP3A4 mRNA in both types of cells. The level of CYP3A4 protein in human hepatocytes was increased by Orlistat after 48 h, while rifampicin strongly induced CYP3A4 protein level. In addition, Orlistat moderately dose-independently activated pregnane X receptor (PXR) in LS174T cells transiently transfected with p3A4-luc reporter construct containing the basal promoter (−362/+53) with proximal PXR response element and the distal xenobiotic responsive enhancer module (−7836/−7208) of the CYP3A4 gene 5′-flanking region. In conclusion, we report here that Orlistat is weak PXR activator and CYP3A4 inducer in human hepatocytes, but it has no effect on CYP3A4 in intestinal cells, implying no role of CYP3A4 induction in the interaction between Orlistat and Cyclosporin in absorption process.